C-3 and C-17 modified triterpenoids as HIV-1 inhibitors

ABSTRACT

Compounds having drug and bio-affecting properties, their pharmaceutical compositions and methods of use are set forth. In particular, betulinic acid derivatives that possess unique antiviral activity are provided as HIV maturation inhibitors, as represented by compounds of Formula I: 
                         
These compounds are useful for the treatment of HIV and AIDS.

FIELD OF THE INVENTION

The present invention relates to novel compounds useful against HIV and,more particularly, to compounds derived from betulinic acid and othercompounds which are useful as HIV maturation inhibitors, and topharmaceutical compositions containing same, as well as to methods fortheir preparation.

BACKGROUND OF THE INVENTION

HIV-1 (human immunodeficiency virus-1) infection remains a major medicalproblem, with an estimated 45-50 million people infected worldwide atthe end of 2010. The number of cases of HIV and AIDS (acquiredimmunodeficiency syndrome) has risen rapidly. In 2005, approximately 5.0million new infections were reported, and 3.1 million people died fromAIDS. Currently available drugs for the treatment of HIV includenucleoside reverse transcriptase (RT) inhibitors or approved single pillcombinations: zidovudine (or AZT or RETROVIR®), didanosine (or VIDEX®),stavudine (or ZERIT®), lamivudine (or 3TC or EPIVIR®), zalcitabine (orDDC or HMD®), abacavir succinate (or ZIAGEN®), tenofovir disoproxilfumarate salt (or VIREAD®), emtricitabine (or FTC-EMTRIVA®), COMBIVIR®(contains −3TC plus AZT), TRIZIVIR® (contains abacavir, lamivudine, andzidovudine), EPZICOM® (contains abacavir and lamivudine), TRUVADA®(contains VIREAD® and EMTRIVA®); non-nucleoside reverse transcriptaseinhibitors: nevirapine (or VIRAMUNE®), delavirdine (or RESCRIPTOR®) andefavirenz (or SUSTIVA®), ATRIPLA® (TRUVADA®+SUSTIVA®), and etravirine,and peptidomimetic protease inhibitors or approved formulations:saquinavir, indinavir, ritonavir, nelfinavir, amprenavir, lopinavir,KALETRA® (lopinavir and Ritonavir), darunavir, atazanavir (REYATAZ®) andtipranavir (APTIVUS®) and cobicistat, and integrase inhibitors such asraltegravir (ISENTRESS®), and entry inhibitors such as enfuvirtide(T-20) (FUZEON®) and maraviroc (SELZENTRY®).

Each of these drugs can only transiently restrain viral replication ifused alone. However, when used in combination, these drugs have aprofound effect on viremia and disease progression. In fact, significantreductions in death rates among AIDS patients have been recentlydocumented as a consequence of the widespread application of combinationtherapy. However, despite these impressive results, 30 to 50% ofpatients may ultimately fail combination drug therapies. Insufficientdrug potency, non-compliance, restricted tissue penetration anddrug-specific limitations within certain cell types (e.g. mostnucleoside analogs cannot be phosphorylated in resting cells) mayaccount for the incomplete suppression of sensitive viruses.Furthermore, the high replication rate and rapid turnover of HIV-1combined with the frequent incorporation of mutations, leads to theappearance of drug-resistant variants and treatment failures whensub-optimal drug concentrations are present. Therefore, novel anti-HIVagents exhibiting distinct resistance patterns, and favorablepharmacokinetic as well as safety profiles are needed to provide moretreatment options. Improved HIV fusion inhibitors and HIV entrycoreceptor antagonists are two examples of new classes of anti-HIVagents further being studied by a number of investigators.

HIV attachment inhibitors are a further subclass of antiviral compoundsthat bind to the HIV surface glycoprotein gp120, and interfere with theinteraction between the surface protein gp120 and the host cell receptorCD4. Thus, they prevent HIV from attaching to the human CD4 T-cell, andblock HIV replication in the first stage of the HIV life cycle. Theproperties of HIV attachment inhibitors have been improved in an effortto obtain compounds with maximized utility and efficacy as antiviralagents. In particular, U.S. Pat. Nos. 7,354,924 and 7,745,625 areillustrative of HIV attachment inhibitors.

Another emerging class of compounds for the treatment of HIV are calledHIV maturation inhibitors. Maturation is the last of as many as 10 ormore steps in HIV replication or the HIV life cycle, in which HIVbecomes infectious as a consequence of several HIV protease-mediatedcleavage events in the gag protein that ultimately results in release ofthe capsid (CA) protein. Maturation inhibitors prevent the HIV capsidfrom properly assembling and maturing, from forming a protective outercoat, or from emerging from human cells. Instead, non-infectious virusesare produced, preventing subsequent cycles of HIV infection.

Certain derivatives of betulinic acid have now been shown to exhibitpotent anti-HIV activity as HIV maturation inhibitors. For example, U.S.Pat. No. 7,365,221 discloses monoacylated betulin and dihydrobetulinederivatives, and their use as anti-HIV agents. As discussed in the '221reference, esterification of betulinic acid (1) with certain substitutedacyl groups, such as 3′,3′-dimethylglutaryl and 3′,3′-dimethylsuccinylgroups produced derivatives having enhanced activity (Kashiwada, Y., etal., J. Med. Chem. 39:1016-1017 (1996)). Acylated betulinic acid anddihydrobetulinic acid derivatives that are potent anti-HIV agents arealso described in U.S. Pat. No. 5,679,828. Esterification of thehydroxyl in the 3 carbon of betulin with succinic acid also produced acompound capable of inhibiting HIV-1 activity (Pokrovskii, A. G., etal., “Synthesis of derivatives of plant triterpenes and study of theirantiviral and immunostimulating activity,” Khimiyay InteresakhUstoichivogo Razvitiya, Vol. 9, No. 3, pp. 485-491 (2001) (Englishabstract).

Other references to the use of treating HIV infection with compoundsderived from betulinic acid include US 2005/0239748 and US 2008/0207573,as well as WO2006/053255, WO2009/100532 and WO2011/007230.

One HIV maturation compound that has been in development has beenidentified as Bevirimat or PA-457, with the chemical formula of C₃₆H₅₆O₆and the IUPAC name of 3β-(3-carboxy-3-methyl-butanoyloxy)lup-20(29)-en-28-oic acid.

Reference is also made herein to the applications by Bristol-MyersSquibb entitled “MODIFIED C-3 BETULINIC ACID DERIVATIVES AS HIVMATURATION INHIBITORS” U.S. Ser. No. 13/151,706 filed on Jun. 2, 2011(now U.S. Pat. No. 8,754,068) and “C-28 AMIDES OF MODIFIED C-3 BETULINICACID DERIVATIVES AS HIV MATURATION INHIBITORS” U.S. Ser. No. 13/151,722,filed on Jun. 2, 2011 (now U.S. Pat. No. 8,802,661). Reference is alsomade to the application entitled “C-28 AMINES OF C-3 MODIFIED BETULINICACID DERIVATIVES AS HIV MATURATION INHIBITORS” U.S. Ser. No. 13/359,680,filed on Jan. 27, 2012 (now U.S. Pat. No. 8,748,415). In addition,reference is made to the application entitled “C-17 AND C-3 MODIFIEDTRITERPENOIDS WITH HIV MATURATION INHIBITORY ACTIVITY” U.S. Ser. No.13/359,727 filed on Jan. 27, 2012 (now U.S. Pat. No. 8,846,647). Furtherreference is also made to the application “C-3 CYCLOALKENYLTRITERPENOIDS WITH HIV MATURATION INHIBITORY ACTIVITY” filed U.S. Ser.No. 13/760,726 on Feb. 6, 2013 (now U.S. Pat. No. 8,906,889), as well asto the application entitled “TRITERPENOIDS WITH HIV MATURATIONINHIBITORY ACTIVITY” U.S. Ser. No. 14/682,179 filed on Apr. 9, 2015.

What is now needed in the art are new compounds which are useful as HIVmaturation inhibitors, as well as new pharmaceutical compositionscontaining these compounds. In particular, new compounds are needed thatwill be effective against emerging genotypic HIV mutants.

SUMMARY OF THE INVENTION

The present invention provides compounds of Formula I below, includingpharmaceutically acceptable salts thereof, their pharmaceuticalformulations, and their use in patients suffering from or susceptible toa virus such as HIV. The compounds of Formula I are effective antiviralagents, particularly as inhibitors of HIV. They are useful for thetreatment of HIV and AIDS.

One embodiment of the present invention is directed to a compound ofFormula I, including pharmaceutically acceptable salts thereof:

wherein R₁ is isopropenyl or isopropyl;A is —C₁₋₆alkyl-OR₀;wherein R₀ is heteroaryl-Q₀;Q₀ is selected from the group of —H, —CN, —C₁₋₆ alkyl, —COOH, -Ph,—OC₁₋₆ alkyl, -halo, —CF₃,Y is selected from the groupof —COOR₂, —C(O)NR₂SO₂R₃, —C(O)NHSO₂NR₂R₂, —SO₂NR₂C(O)R₂, -tetrazole,and —CONHOH;R₂ is —H, —C₁₋₆ alkyl, -alkylsubstituted C₁₋₆ alkyl or -arylsubstitutedC₁₋₆ alkyl;W is absent, or is —CH₂— or —CO—;R₃ is —H, —C₁₋₆ alkyl or -alkylsubstituted C₁₋₆ alkyl;R₄ is selected from the group of —H, —C₁₋₆ alkyl, —C₁₋₆ alky 1-C₃₋₆cycloalkyl, —C₁₋₆ substituted —C₁₋₆ alkyl, —C₁₋₆ alkyl-Q₁,—C₁₋₆alkyl-C₃₋₆ cycloalkyl-Q₁, aryl, heteroaryl, substituted heteroaryl,—COR₆, —SO₂R₇, —SO₂NR₂R₂, and

wherein G is selected from the group of —O—, —SO₂— and —NR₁₂—;wherein Q₁ is selected from the group of —C₁₋₆ alkyl, —C₁₋₆ fluoroalkyl,heteroaryl, substituted heteroaryl, halogen, —CF₃, —OR₂, —COOR₂, —NR₈R₉,—CONR₈R₉ and —SO₂R₇;R₅ is selected from the group of —H, —C₁₋₆ alkyl, —C₃₋₆ cycloalkyl,—C₁₋₆ alkylsubstituted alkyl, —C₁₋₆ alkyl-NR₈R₉, —COR₃, —SO₂R₇ and—SO₂NR₂R₂;with the proviso that R₄ or R₅ is not —COR₆ when W is —CO—;with the further proviso that only one of R₄ or R₅ is selected from thegroup of —COR₆, —COCOR₆, —SO₂R₇ and —SO₂NR₂R₂;R₆ is selected from the group of —H, —C₁₋₆ alkyl, —C₁₋₆alkyl-substituted alkyl, —C₃₋₆ cycloalkyl, —C₃₋₆ substitutedcycloalkyl-Q₂, —C₁₋₆alkyl-Q₂, —C₁₋₆ alkyl-substituted alkyl-Q₂, —C₃₋₆cycloalkyl-Q₂, aryl-Q₂, —NR₁₃R₁₄, and —OR₁₅;wherein Q₂ is selected from the group of aryl, heteroaryl, substitutedheteroaryl, —OR₂, —COOR₂, —NR₈R₉, SO₂R₇, —CONHSO₂R₃, and —CONHSO₂NR₂R₂;R₇ is selected from the group of —H, —C₁₋₆ alkyl, —C₁₋₆ substitutedalkyl, —C₃₋₆ cycloalkyl, —CF₃, aryl, and heteroaryl;R₈ and R₉ are independently selected from the group of —H, —C₁₋₆ alkyl,—C₁₋₆ substituted alkyl, aryl, heteroaryl, substituted aryl, substitutedheteroaryl, —C₁₋₆ alkyl-Q₂, and —COOR₃, or R⁸ and R₉ are taken togetherwith the adjacent N to form a cycle selected from the group of:

M is selected from the group of —R₁₅, —SO₂R₂, —SO₂NR₂R₂, —OH and—NR₂R₁₂;V is selected from the group of —CR₁₀R₁₁—, —SO₂—, —O— and —NR₁₂—;with the proviso that only one of R₈ or R₉ can be —COOR₃;R₁₀ and Rn are independently selected from the group of —H, —C₁₋₆ alkyl,—C₁₋₆ substituted alkyl and —C₃₋₆ cycloalkyl;R₁₂ is selected from the group of —H, —C₁₋₆ alkyl, -alkylsubstitutedC₁₋₆ alkyl, —CONR₂R₂, —SO₂R₃, and —SO₂NR₂R₂;R₁₃ and R₁₄ are independently selected from the group of —H, —C₁₋₆alkyl, —C₃₋₆ cycloalkyl, —C₁₋₆ substituted alkyl, —C₁₋₆ alkyl-Q₃, —C₁₋₆alkyl-C₃₋₆ cycloalkyl-Q₃, and C₁₋₆ substituted alkyl-Q₃;Q₃ is selected from the group of heteroaryl, substituted heteroaryl,—NR₂R₁₂, —CONR₂R₂, —COOR₂, —OR₂, and —SO₂R₃;R₁₅ is selected from the group of —C₁₋₆ alkyl, —C₃₋₆ cycloalkyl, —C₁₋₆substituted alkyl, —C₁₋₆ alkyl-Q₃, —C₁₋₆ alkyl-C₃₋₆ cycloalkyl-Q₃ and—C₁₋₆ substituted alkyl-Q₃;R₁₆ is selected from the group of —H, —C₁₋₆ alkyl, —NR₂R₂, and —COOR₂;with the proviso that when V is —NR₁₂—; R₁₆ is not —NR₂R₂; andR₁₇ is selected from the group of —H, —C₁₋₆ alkyl, —COOR₃, and aryl.

In a further embodiment, there is provided a method for treating mammalsinfected with a virus, especially wherein said virus is HIV, comprisingadministering to said mammal an antiviral effective amount of a compoundwhich is selected from the group of compounds of Formula I, and one ormore pharmaceutically acceptable carriers, excipients or diluents.Optionally, the compound of Formula I can be administered in combinationwith an antiviral effective amount of another AIDS treatment agentselected from the group consisting of: (a) an AIDS antiviral agent; (b)an anti-infective agent; (c) an immunomodulator; and (d) other HIV entryinhibitors.

Another embodiment of the present invention is a pharmaceuticalcomposition comprising one or more compounds of Formula I, and one ormore pharmaceutically acceptable carriers, excipients, and/or diluents;and optionally in combination with another AIDS treatment agent selectedfrom the group consisting of: (a) an AIDS antiviral agent; (b) ananti-infective agent; (c) an immunomodulator; and (d) other HIV entryinhibitors.

In another embodiment of the invention there is provided one or moremethods for making the compounds of Formula I herein.

Also provided herein are intermediate compounds useful in making thecompounds of Formula I herein.

The present invention is directed to these, as well as other importantends, hereinafter described.

DETAILED DESCRIPTION OF THE EMBODIMENTS

As used herein, the singular forms “a”, “an”, and “the” include pluralreference unless the context clearly dictates otherwise.

Since the compounds of the present invention may possess asymmetriccenters and therefore occur as mixtures of diastereomers, the presentdisclosure includes the individual diastereoisomeric forms of thecompounds of Formula I in addition to the mixtures thereof.

Definitions

Unless otherwise specifically set forth elsewhere in the application,one or more of the following terms may be used herein, and shall havethe following meanings:

“H” refers to hydrogen, including its isotopes, such as deuterium.

The term “C₁₋₆ alkyl” as used herein and in the claims (unless specifiedotherwise) mean straight or branched chain alkyl groups such as methyl,ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, amyl, hexyl and thelike.

“C₁-C₄ fluoroalkyl” refers to F-substituted —C₄ alkyl wherein at leastone H atom is substituted with F atom, and each H atom can beindependently substituted by F atom;

“Halogen” or “halo” refers to chlorine, bromine, iodine or fluorine.

An “aryl” or “Ar” group refers to an all carbon monocyclic or fused-ringpolycyclic (i.e., rings which share adjacent pairs of carbon atoms)groups having a completely conjugated pi-electron system. Examples,without limitation, of aryl groups are phenyl, naphthalenyl andanthracenyl. The aryl group may be substituted or unsubstituted. Whensubstituted, the substituent group(s) are preferably one or moreselected from alkyl, cycloalkyl, aryl, heteroaryl, heteroalicyclic,hydroxy, alkoxy, aryloxy, heteroaryloxy, heteroalicycloxy, thiohydroxy,thioaryloxy, thioheteroaryloxy, thioheteroalicycloxy, cyano, halogen,nitro, carbonyl, O-carbamyl, N-carbamyl, C-amido, N-amido, C-carboxy,O-carboxy, sulfinyl, sulfonyl, sulfonamido, trihalomethyl, ureido, aminoand —NR^(x)R^(y), wherein R^(x) and R^(y) are independently selectedfrom the group consisting of hydrogen, alkyl, cycloalkyl, aryl,carbonyl, C-carboxy, sulfonyl, trihalomethyl, and, combined, a five- orsix-member heteroalicyclic ring.

A “heteroaryl” group refers to a monocyclic or fused ring (i.e., ringswhich share an adjacent pair of atoms) group having in the ring(s) oneor more atoms selected from the group consisting of nitrogen, oxygen andsulfur and, in addition, having a completely conjugated pi-electronsystem. Unless otherwise indicated, the heteroaryl group may be attachedat either a carbon or nitrogen atom within the heteroaryl group. Itshould be noted that the term heteroaryl is intended to encompass anN-oxide of the parent heteroaryl if such an N-oxide is chemicallyfeasible as is known in the art. Examples, without limitation, ofheteroaryl groups are furyl, thienyl, benzothienyl, thiazolyl,imidazolyl, oxazolyl, oxadiazolyl, thiadiazolyl, benzothiazolyl,triazolyl, tetrazolyl, isoxazolyl, isothiazolyl, pyrrolyl, pyranyl,tetrahydropyranyl, pyrazolyl, pyridyl, pyrimidinyl, quinolinyl,isoquinolinyl, purinyl, carbazolyl, benzoxazolyl, benzimidazolyl,indolyl, isoindolyl, pyrazinyl, diazinyl, pyrazine, triazinyl,tetrazinyl, and tetrazolyl. When substituted the substituted group(s) ispreferably one or more selected from alkyl, cycloalkyl, aryl,heteroaryl, heteroalicyclic, hydroxy, alkoxy, aryloxy, heteroaryloxy,heteroalicycloxy, thioalkoxy, thiohydroxy, thioaryloxy,thioheteroaryloxy, thioheteroalicycloxy, cyano, halogen, nitro,carbonyl, O-carbamyl, N-carbamyl, C-amido, N-amido, C-carboxy,O-carboxy, sulfinyl, sulfonyl, sulfonamido, trihalomethyl, ureido,amino, and —NR^(x)R^(y), wherein R^(x) and R^(y) are as defined above.

A “heteroalicyclic” group refers to a monocyclic or fused ring grouphaving in the ring(s) one or more atoms selected from the groupconsisting of nitrogen, oxygen and sulfur. Rings are selected from thosewhich provide stable arrangements of bonds and are not intended toencompass systems which would not exist. The rings may also have one ormore double bonds. However, the rings do not have a completelyconjugated pi-electron system. Examples, without limitation, ofheteroalicyclic groups are azetidinyl, piperidyl, piperazinyl,imidazolinyl, thiazolidinyl, 3-pyrrolidin-1-yl, morpholinyl,thiomorpholinyl and its S oxides and tetrahydropyranyl. When substitutedthe substituted group(s) is preferably one or more selected from alkyl,cycloalkyl, aryl, heteroaryl, heteroalicyclic, hydroxy, alkoxy, aryloxy,heteroaryloxy, heteroalicycloxy, thiohydroxy, thioalkoxy, thioaryloxy,thioheteroaryloxy, thioheteroalicycloxy, cyano, halogen, nitro,carbonyl, thiocarbonyl, O-carbamyl, N-carbamyl, O-thiocarbamyl,N-thiocarbamyl, C-amido, C-thioamido, N-amido, C-carboxy, O-carboxy,sulfinyl, sulfonyl, sulfonamido, trihalomethanesulfonamido,trihalomethanesulfonyl, silyl, guanyl, guanidino, ureido, phosphonyl,amino and —NR^(x)R^(y), wherein R^(x) and R^(y) are as defined above.

An “alkyl” group refers to a saturated aliphatic hydrocarbon includingstraight chain and branched chain groups. Preferably, the alkyl grouphas 1 to 20 carbon atoms (whenever a numerical range; e.g., “1-20”, isstated herein, it means that the group, in this case the alkyl group maycontain 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc. up to andincluding 20 carbon atoms). More preferably, it is a medium size alkylhaving 1 to 10 carbon atoms. Most preferably, it is a lower alkyl having1 to 4 carbon atoms. The alkyl group may be substituted orunsubstituted. When substituted, the substituent group(s) is preferablyone or more individually selected from trihaloalkyl, cycloalkyl, aryl,heteroaryl, heteroalicyclic, hydroxy, alkoxy, aryloxy, heteroaryloxy,heteroalicycloxy, thiohydroxy, thioalkoxy, thioaryloxy,thioheteroaryloxy, thioheteroalicycloxy, cyano, halo, nitro, carbonyl,thiocarbonyl, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl,C-amido, C-thioamido, N-amido, C-carboxy, O-carboxy, sulfinyl, sulfonyl,sulfonamido, trihalomethanesulfonamido, trihalomethanesulfonyl, andcombined, a five- or six-member heteroalicyclic ring.

A “cycloalkyl” group refers to an all-carbon monocyclic or fused ring(i.e., rings which share and adjacent pair of carbon atoms) groupwherein one or more rings does not have a completely conjugatedpi-electron system. Examples, without limitation, of cycloalkyl groupsare cyclopropane, cyclobutane, cyclopentane, cyclopentene, cyclohexane,cyclohexene, cycloheptane, cycloheptene and adamantane. A cycloalkylgroup may be substituted or unsubstituted. When substituted, thesubstituent group(s) is preferably one or more individually selectedfrom alkyl, aryl, heteroaryl, heteroalicyclic, hydroxy, alkoxy, aryloxy,heteroaryl oxy, heteroalicycloxy, thiohydroxy, thioalkoxy, thioaryloxy,thioheteroaryloxy, thioheteroalicycloxy, cyano, halo, nitro, carbonyl,thiocarbonyl, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl,C-amido, C-thioamido, N-amido, C-carboxy, O-carboxy, sulfinyl, sulfonyl,sulfonamido, trihalomethanesulfonamido, trihalomethanesulfonyl, silyl,amidino, guanidino, ureido, phosphonyl, amino and —NR^(x)R^(y) withR^(x) and R^(y) as defined above.

An “alkenyl” group refers to an alkyl group, as defined herein, havingat least two carbon atoms and at least one carbon-carbon double bond.

An “alkynyl” group refers to an alkyl group, as defined herein, havingat least two carbon atoms and at least one carbon-carbon triple bond.

A “hydroxy” group refers to an —OH group.

An “alkoxy” group refers to both an —O-alkyl and an —O-cycloalkyl groupas defined herein.

An “aryloxy” group refers to both an —O-aryl and an —O-heteroaryl group,as defined herein.

A “heteroaryloxy” group refers to a heteroaryl-O— group with heteroarylas defined herein.

A “heteroalicycloxy” group refers to a heteroalicyclic-O— group withheteroalicyclic as defined herein.

A “thiohydroxy” group refers to an —SH group.

A “thioalkoxy” group refers to both an S-alkyl and an —S-cycloalkylgroup, as defined herein.

A “thioaryloxy” group refers to both an —S-aryl and an —S-heteroarylgroup, as defined herein.

A “thioheteroaryloxy” group refers to a heteroaryl-S— group withheteroaryl as defined herein.

A “thioheteroalicycloxy” group refers to a heteroalicyclic-S— group withheteroalicyclic as defined herein.

A “carbonyl” group refers to a —C(═O)—R″ group, where R″ is selectedfrom the group consisting of hydrogen, alkyl, alkenyl, alkynyl,cycloalkyl, aryl, heteroaryl (bonded through a ring carbon) andheteroalicyclic (bonded through a ring carbon), as each is definedherein.

An “aldehyde” group refers to a carbonyl group where R″ is hydrogen.

A “thiocarbonyl” group refers to a —C(═S)—R″ group, with R″ as definedherein.

A “keto” group refers to a —CC(═O)C— group wherein the carbon on eitheror both sides of the C═O may be alkyl, cycloalkyl, aryl or a carbon of aheteroaryl or heteroalicyclic group.

A “trihalomethanecarbonyl” group refers to a Z₃CC(═O)— group with said Zbeing a halogen.

A “C-carboxy” group refers to a —C(═O)O—R″ groups, with R″ as definedherein.

An “O-carboxy” group refers to a R″C(═O)O— group, with R″ as definedherein.

A “carboxylic acid” group refers to a C-carboxy group in which R″ ishydrogen.

A “trihalomethyl” group refers to a —CZ₃, group wherein Z is a halogengroup as defined herein.

A “trihalomethanesulfonyl” group refers to a Z₃CS(═O)₂— groups with Z asdefined above.

A “trihalomethanesulfonamido” group refers to a Z₃CS(═O)₂NR^(x)— groupwith Z as defined above and R^(x) being H or (C₁₋₆)alkyl.

A “sulfinyl” group refers to a —S(═O)—R″ group, with R″ being(C₁₋₆)alkyl.

A “sulfonyl” group refers to a —S(═O)₂R″ group with R″ being(C₁₋₆)alkyl.

A “S-sulfonamido” group refers to a —S(═O)₂NR^(X)R^(Y), with R^(X) andR^(Y) independently being H or (C₁₋₆)alkyl.

A “N-sulfonamido” group refers to a R″S(═O)₂NR_(X)— group, with R_(x)being H or (C₁₋₆)alkyl.

A “O-carbamyl” group refers to a —OC(═O)NR^(x)R^(y) group, with R^(X)and R^(Y) independently being H or (C₁₋₆)alkyl.

A “N-carbamyl” group refers to a R^(x)OC(═O)NR^(y) group, with R^(x) andR^(y) independently being H or (C₁₋₆)alkyl.

A “O-thiocarbamyl” group refers to a —OC(═S)NR^(x)R^(y) group, withR^(x) and R^(y) independently being H or (C₁₋₆)alkyl.

A “N-thiocarbamyl” group refers to a R^(x)OC(═S)NR^(y)— group, withR^(x) and R^(y) independently being H or (C₁₋₆)alkyl.

An “amino” group refers to an —NH₂ group.

A “C-amido” group refers to a —C(═O)NR^(x)R^(y) group, with R^(x) andR^(y) independently being H or (C₁₋₆)alkyl.

A “C-thioamido” group refers to a —C(═S)NR^(x)R^(y) group, with R^(x)and R^(y) independently being H or (C₁₋₆)alkyl.

A “N-amido” group refers to a R^(x)C(═O)NR^(y)— group, with R^(x) andR^(y) independently being H or (C₁₋₆)alkyl.

An “ureido” group refers to a —NR^(x)C(═O)NR^(y)R^(y2) group, withR^(x), R^(y), and R^(y2) independently being H or (C₁₋₆)alkyl.

A “guanidino” group refers to a —R^(x)NC(═N)NR^(y)R^(y2) group, withR^(x), R^(y), and R^(y2) independently being H or (C₁₋₆)alkyl.

A “amidino” group refers to a R^(x)R^(y)NC(═N)— group, with R^(x) andR^(y) independently being H or (C₁₋₆)alkyl.

A “cyano” group refers to a —CN group.

A “silyl” group refers to a —Si(R″)₃, with R″ being (C₁₋₆)alkyl orphenyl.

A “phosphonyl” group refers to a P(═O)(OR^(x))₂ with R^(x) being(C₁₋₆)alkyl.

A “hydrazino” group refers to a —NR^(x)NR^(y)R^(y2) group, with R^(x),R^(y), and R^(y2) independently being H or (C₁₋₆)alkyl.

A “4, 5, or 6 membered ring cyclic N-lactam” group refers to

A “spiro” group is a bicyclic organic group with rings connected throughjust one atom. The rings can be different in nature or identical. Theconnecting atom is also called the spiroatom, most often a quaternarycarbon (“spiro carbon”).

An “oxospiro” or “oxaspiro” group is a spiro group having an oxygencontained within the bicyclic ring structure. A “dioxospiro” or“dioxaspiro” group has two oxygens within the bicyclic ring structure.

Any two adjacent R groups may combine to form an additional aryl,cycloalkyl, heteroaryl or heterocyclic ring fused to the ring initiallybearing those R groups.

It is known in the art that nitrogen atoms in heteroaryl systems can be“participating in a heteroaryl ring double bond”, and this refers to theform of double bonds in the two tautomeric structures which comprisefive-member ring heteroaryl groups. This dictates whether nitrogens canbe substituted as well understood by chemists in the art. The disclosureand claims of the present disclosure are based on the known generalprinciples of chemical bonding. It is understood that the claims do notencompass structures known to be unstable or not able to exist based onthe literature.

Pharmaceutically acceptable salts and prodrugs of compounds disclosedherein are within the scope of the invention. The term “pharmaceuticallyacceptable salt” as used herein and in the claims is intended to includenontoxic base addition salts. Suitable salts include those derived fromorganic and inorganic acids such as, without limitation, hydrochloricacid, hydrobromic acid, phosphoric acid, sulfuric acid, methanesulfonicacid, acetic acid, tartaric acid, lactic acid, sulfinic acid, citricacid, maleic acid, fumaric acid, sorbic acid, aconitic acid, salicylicacid, phthalic acid, and the like. The term “pharmaceutically acceptablesalt” as used herein is also intended to include salts of acidic groups,such as a carboxylate, with such counterions as aminonium, alkali metalsalts, particularly sodium or potassium, alkaline earth metal salts,particularly calcium or magnesium, and salts with suitable organic basessuch as lower alkylamines (methylamine, ethylamine, cyclohexylamine, andthe like) or with substituted lower alkylamines (e.g.hydroxyl-substituted alkylamines such as diethanolamine, triethanolamineor tris(hydroxymethyl)-aminomethane), or with bases such as piperidineor morpholine.

As stated above, the compounds of the invention also include “prodrugs”.The term “prodrug” as used herein encompasses both the term “prodrugesters” and the term “prodrug ethers”.

As set forth above, the invention is directed to a compound, includingpharmaceutically acceptable salts thereof, which is selected from acompound of Formula

wherein R₁ is isopropenyl or isopropyl;A is —C₁₋₆ alkyl-OR₀;wherein R₀ is heteroaryl-Q₀;Q₀ is selected from the group of —H, —CN, —C₁₋₆ alkyl, —COOH, -Ph,—OC₁₋₆ alkyl, -halo, —CF₃,Y is selected from the group of —COOR₂, —C(O)NR₂SO₂R₃, —C(O)NHSO₂NR₂R₂,—SO₂NR₂C(O)R₂, -tetrazole, and —CONHOH;R₂ is —H, —C₁₋₆ alkyl, -alkylsubstituted C₁₋₆ alkyl or -arylsubstitutedC₁₋₆ alkyl;W is absent, or is —CH2- or —CO—;R₃ is —H, —C₁₋₆ alkyl or -alkylsubstituted C₁₋₆ alkyl;R₄ is selected from the group of —H, —C₁₋₆ alkyl, —C₁₋₆alkyl-C₃₋₆cycloalkyl, —C₁₋₆ substituted —C₁₋₆ alkyl, —C₁₋₆ alkyl-Q₁, —C₁₋₆alkyl-C₃₋₆ cycloalkyl-Q₁, aryl, heteroaryl, substituted heteroaryl,—COR₆, —SO₂R₇, —SO₂NR₂R₂, and

wherein G is selected from the group of —O—, —SO₂— and —NR₁₂—;wherein Q₁ is selected from the group of —C₁₋₆ alkyl, —C₁₋₆ fluoroalkyl,heteroaryl, substituted heteroaryl, halogen, —CF₃, —OR₂, —COOR₂, —NR₈R₉,—CONR₈R₉ and —SO₂R₇;R₅ is selected from the group of —H, —C₁₋₆ alkyl, —C₃₋₆ cycloalkyl,—C₁₋₆ alkylsubstituted alkyl, —C₁₋₆ alkyl-NR₈R₉, —COR₃, —SO₂R₇ and—SO₂NR₂R₂;with the proviso that R₄ or R₅ is not —COR₆ when W is —CO—;with the further proviso that only one of R₄ or R₅ is selected from thegroup of —COR₆, —COCOR₆, —SO₂R₇ and —SO₂NR₂R₂;R₆ is selected from the group of —H, —C₁₋₆ alkyl, —C₁₋₆alkyl-substitutedalkyl, —C₃₋₆ cycloalkyl, —C₃₋₆substitutedcycloalkyl-Q₂, —C₁₋₆ alkyl-Q₂, —C₁₋₆alkyl-substitutedalkyl-Q₂, —C₃₋₆ cycloalkyl-Q₂, aryl-Q₂, —NR₁₃R₁₄, and—OR₁₅;wherein Q₂ is selected from the group of aryl, heteroaryl, substitutedheteroaryl, —OR₂, —COOR₂, —NR₈R₉, SO₂R₇, —CONHSO₂R₃, and —CONHSO₂NR₂R₂;R₇ is selected from the group of —H, —C₁₋₆ alkyl, —C₁₋₆ substitutedalkyl, —C₃₋₆ cycloalkyl, —CF₃, aryl, and heteroaryl;R₈ and R₉ are independently selected from the group of —H, —C₁₋₆ alkyl,—C₁₋₆ substituted alkyl, aryl, heteroaryl, substituted aryl, substitutedheteroaryl, —C₁₋₆ alkyl-Q₂, and —COOR₃, or R₈ and R₉ are taken togetherwith the adjacent N to form a cycle selected from the group of:

M is selected from the group of —R₁₅, —SO₂R₂, —SO₂NR₂R₂, —OH and—NR₂R₁₂;V is selected from the group of —CR₁₀R₁₁—, —SO₂—, —O— and —NR₁₂—;with the proviso that only one of R₈ or R₉ can be —COOR₃;R₁₀ and R₁₁ are independently selected from the group of —H, —C₁₋₆alkyl, —C₁₋₆ substituted alkyl and —C₃₋₆ cycloalkyl;R₁₂ is selected from the group of —H, —C₁₋₆ alkyl, -alkylsubstitutedC₁₋₆ alkyl, —CONR₂R₂, —SO₂R₃, and —SO₂NR₂R₂;R₁₃ and RH are independently selected from the group of —H, —C₁₋₆ alkyl,—C₃₋₆ cycloalkyl, —C₁₋₆ substituted alkyl, —C₁₋₆alkyl-Q₃,—C₁₋₆alkyl-C₃₋₆ cycloalkyl-Q₃, and C₁₋₆ substituted alkyl-Q₃;Q₃ is selected from the group of heteroaryl, substituted heteroaryl,—NR₂R₁₂, —CONR₂R₂, —COOR₂, —OR₂, and —SO₂R₃;R₁₅ is selected from the group of —C₁₋₆ alkyl, —C₃₋₆ cycloalkyl, —C₁₋₆substituted alkyl, —C₁₋₆ alkyl-Q₃, —C₁₋₆ alkyl-C₆₋₆ cycloalkyl-Q₃ and—C₁₋₆ substituted alkyl-Q₃;R₁₆ is selected from the group of —H, —C₁₋₆ alkyl, —NR₂R₂, and —COOR₂;with the proviso that when V is —NR₁₂—; R₁₆ is not —NR₂R₂; andR₁₇ is selected from the group of —H, —C₁₋₆ alkyl, —COOR₃, and aryl.

In a preferred embodiment of the invention, R₁ is isopropenyl.

It is also preferred that Y is —COOR₂. More preferably, R₂ in thisembodiment is —H.

In another preferred embodiment of the invention, in the R₀ group the“heteroaryl” moiety is preferably selected from the group of:

It is also preferred that there is no intervening alkyl group or othersubstituent group between the —O moiety and the R₀ group in substituentA.

It is further preferred that R₄ is —C₁₋₆ alkyl-Q₁.

Also preferred is the embodiment wherein Q₁ is —NR₈R₉.

Additionally, when R₈ and R₉ are taken together with the adjacent —N toform a cycle, the preferred cycle will be selected from the group of:

In some embodiments it is also preferred that Q₀ is —CN.

In another preferred embodiment, R₁ is isopropenyl, in the R₀ group the“heteroaryl” moiety is selected from the group of:

Y is —COOH, R₄ is —C₁₋₆ alkyl-Qt, Q₁ is —NR₈R₉, and R₈ and R₉ are takentogether with the adjacent —N to form a cycle which is selected from thegroup of:

In this embodiment, it is also preferred that R₇ and R₁₆ are each —H or—C₁₋₆ alkyl.

Preferred compounds, including pharmaceutically acceptable saltsthereof, as part of the invention include the following:

In another embodiment, preferred compounds, including pharmaceuticallyacceptable salts thereof, will be the following:

The compounds above represent the mixture of diastereoisomers, and thetwo individual diastereomers. In certain embodiments, one of thespecific diastereomers may be particularly preferred.

The compounds of the present invention, according to all the variousembodiments described above, may be administered orally, parenterally(including subcutaneous injections, intravenous, intramuscular,intrasternal injection or infusion techniques), by inhalation spray, orrectally, and by other means, in dosage unit formulations containingnon-toxic pharmaceutically acceptable carriers, excipients and diluentsavailable to the skilled artisan. One or more adjuvants may also beincluded.

Thus, in accordance with the present invention, there is furtherprovided a method of treatment, and a pharmaceutical composition, fortreating viral infections such as HIV infection and AIDS. The treatmentinvolves administering to a patient in need of such treatment apharmaceutical composition which contains an antiviral effective amountof one or more of the compounds of Formula I together with one or morepharmaceutically acceptable carriers, excipients or diluents. As usedherein, the term “antiviral effective amount” means the total amount ofeach active component of the composition and method that is sufficientto show a meaningful patient benefit, i.e., inhibiting, ameliorating, orhealing of acute conditions characterized by inhibition of HIVinfection. When applied to an individual active ingredient, administeredalone, the term refers to that ingredient alone. When applied to acombination, the term refers to combined amounts of the activeingredients that result in the therapeutic effect, whether administeredin combination, serially or simultaneously. The terms “treat, treating,treatment” as used herein and in the claims means preventing,inhibiting, ameliorating and/or healing diseases and conditionsassociated with HIV infection.

The pharmaceutical compositions of the invention may be in the form oforally administrable suspensions or tablets; as well as nasal sprays,sterile injectable preparations, for example, as sterile injectableaqueous or oleaginous suspensions or suppositories. Pharmaceuticallyacceptable carriers, excipients or diluents may be utilized in thepharmaceutical compositions, and are those utilized in the art ofpharmaceutical preparations.

When administered orally as a suspension, these compositions areprepared according to techniques typically known in the art ofpharmaceutical formulation and may contain microcrystalline cellulosefor imparting bulk, alginic acid or sodium alginate as a suspendingagent, methylcellulose as a viscosity enhancer, and sweeteners/flavoringagents known in the art. As immediate release tablets, thesecompositions may contain microcrystalline cellulose, dicalciumphosphate, starch, magnesium stearate and lactose and/or otherexcipients, binders, extenders, disintegrants, diluents, and lubricantsknown in the art.

The injectable solutions or suspensions may be formulated according toknown art, using suitable non-toxic, parenterally acceptable diluents orsolvents, such as mannitol, 1,3-butanediol, water, Ringer's solution orisotonic sodium chloride solution, or suitable dispersing or wetting andsuspending agents, such as sterile, bland, fixed oils, includingsynthetic mono- or diglycerides, and fatty acids, including oleic acid.

The compounds herein set forth can be administered orally to humans in adosage range of about 1 to 100 mg/kg body weight in divided doses,usually over an extended period, such as days, weeks, months, or evenyears. One preferred dosage range is about 1 to 10 mg/kg body weightorally in divided doses. Another preferred dosage range is about 1 to 20mg/kg body weight in divided doses. It will be understood, however, thatthe specific dose level and frequency of dosage for any particularpatient may be varied and will depend upon a variety of factorsincluding the activity of the specific compound employed, the metabolicstability and length of action of that compound, the age, body weight,general health, sex, diet, mode and time of administration, rate ofexcretion, drug combination, the severity of the particular condition,and the host undergoing therapy.

Also contemplated herein are combinations of the compounds of Formula Iherein set forth, together with one or more other agents useful in thetreatment of AIDS. For example, the compounds of this disclosure may beeffectively administered, whether at periods of pre-exposure and/orpost-exposure, in combination with effective amounts of the AIDSantivirals, immunomodulators, antiinfectives, or vaccines, such as thosein the following non-limiting table:

ANTIVIRALS Drug Name Manufacturer Indication 097 Hoechst/Bayer HIVinfection, AIDS, ARC (non-nucleoside reverse transcriptase (RT)inhibitor) Amprenavir Glaxo Wellcome HIV infection, 141 W94 AIDS, ARC GW141 (protease inhibitor) Abacavir (1592U89) Glaxo Wellcome HIVinfection, GW 1592 AIDS, ARC (RT inhibitor) Acemannan Carrington LabsARC (Irving, TX) Acyclovir Burroughs Wellcome HIV infection, AIDS, ARCAD-439 Tanox Biosystems HIV infection, AIDS, ARC AD-519 Tanox BiosystemsHIV infection, AIDS, ARC Adefovir dipivoxil Gilead Sciences HIVinfection AL-721 Ethigen ARC, PGL (Los Angeles, CA) HIV positive, AIDSAlpha Interferon Glaxo Wellcome Kaposi's sarcoma, HIV in combinationw/Retrovir Ansamycin Adria Laboratories ARC LM 427 (Dublin, OH) Erbamont(Stamford, CT) Antibody which Advanced Biotherapy AIDS, ARC NeutralizespH Concepts Labile alpha aberrant (Rockville, MD) Interferon AR177Aronex Pharm HIV infection, AIDS, ARC Beta-fluoro-ddA Nat'l CancerInstitute AIDS-associated diseases BMS-234475 Bristol-Myers Squibb/ HIVinfection, (CGP-61755) Novartis AIDS, ARC (protease inhibitor) CI-1012Warner-Lambert HIV-1 infection Cidofovir Gilead Science CMV retinitis,herpes, papillomavirus Curdlan sulfate AJI Pharma USA HIV infectionCytomegalovirus MedImmune CMV retinitis Immune globin Cytovene SyntexSight threatening Ganciclovir CMV peripheral CMV retinitis DarunavirTibotec-J & J HIV infection, AIDS, ARC (protease inhibitor) DelaviridinePharmacia-Upjohn HIV infection, AIDS, ARC (RT inhibitor) Dextran SulfateUeno Fine Chem. AIDS, ARC, HIV Ind. Ltd. (Osaka, positive Japan)asymptomatic ddC Hoffman-La Roche HIV infection, AIDS, DideoxycytidineARC ddI Bristol-Myers Squibb HIV infection, AIDS, Dideoxyinosine ARC;combination with AZT/d4T DMP-450 AVID HIV infection, (Camden, NJ) AIDS,ARC (protease inhibitor) Efavirenz Bristol Myers Squibb HIV infection,(DMP 266, SUSTIVA ®) AIDS, ARC (−)6-Chloro-4-(S)- (non-nucleoside RTcyclopropylethynyl- inhibitor) 4(S)-trifluoro- methyl-1,4-dihydro-2H-3,1-benzoxazin- 2-one, STOCRINE EL10 Elan Corp, PLC HIV infection(Gainesville, GA) Etravirine Tibotec/J & J HIV infection, AIDS, ARC(non-nucleoside reverse transcriptase inhibitor) Famciclovir Smith Klineherpes zoster, herpes simplex GS 840 Gilead HIV infection, AIDS, ARC(reverse transcriptase inhibitor) HBY097 Hoechst Marion HIV infection,Roussel AIDS, ARC (non-nucleoside reverse transcriptase inhibitor)Hypericin VIMRx Pharm. HIV infection, AIDS, ARC Recombinant Human TritonBiosciences AIDS, Kaposi's Interferon Beta (Almeda, CA) sarcoma, ARCInterferon alfa-n3 Interferon Sciences ARC, AIDS Indinavir Merck HIVinfection, AIDS, ARC, asymptomatic HIV positive, also in combinationwith AZT/ddI/ddC ISIS 2922 ISIS Pharmaceuticals CMV retinitis KNI-272Nat'l Cancer Institute HIV-assoc. diseases Lamivudine, 3TC GlaxoWellcome HIV infection, AIDS, ARC (reverse transcriptase inhibitor);also with AZT Lobucavir Bristol-Myers Squibb CMV infection NelfinavirAgouron HIV infection, Pharmaceuticals AIDS, ARC (protease inhibitor)Nevirapine Boeheringer HIV infection, Ingleheim AIDS, ARC (RT inhibitor)Novapren Novaferon Labs, Inc. HIV inhibitor (Akron, OH) Peptide TPeninsula Labs AIDS Octapeptide (Belmont, CA) Sequence Trisodium AstraPharm. CMV retinitis, HIV Phosphonoformate Products, Inc. infection,other CMV infections PNU-140690 Pharmacia Upjohn HIV infection, AIDS,ARC (protease inhibitor) Probucol Vyrex HIV infection, AIDS RBC-CD4Sheffield Med. HIV infection, Tech (Houston, TX) AIDS, ARC RitonavirAbbott HIV infection, AIDS, ARC (protease inhibitor) SaquinavirHoffmann- HIV infection, LaRoche AIDS, ARC (protease inhibitor)Stavudine; d4T Bristol-Myers Squibb HIV infection, AIDS, Didehydrodeoxy-ARC Thymidine Tipranavir Boehringer Ingelheim HIV infection, AIDS, ARC(protease inhibitor) Valaciclovir Glaxo Wellcome Genital HSV & CMVinfections Virazole Viratek/ICN asymptomatic HIV Ribavirin (Costa Mesa,CA) positive, LAS, ARC VX-478 Vertex HIV infection, AIDS, ARCZalcitabine Hoffmann-LaRoche HIV infection, AIDS, ARC, with AZTZidovudine; AZT Glaxo Wellcome HIV infection, AIDS, ARC, Kaposi'ssarcoma, in combination with other therapies Tenofovir disoproxil,Gilead HIV infection, fumarate salt (VIREAD ®) AIDS, (reversetranscriptase inhibitor) EMTRIVA ® Gilead HIV infection, (Emtricitabine)(FTC) AIDS, (reverse transcriptase inhibitor) COMBIVIR ® GSK HIVinfection, AIDS, (reverse transcriptase inhibitor) Abacavir succinateGSK HIV infection, (or ZIAGEN ®) AIDS, (reverse transcriptase inhibitor)REYATAZ ® Bristol-Myers Squibb HIV infection (or atazanavir) AIDs,protease inhibitor FUZEON ® Roche/Trimeris HIV infection (Enfuvirtide orT-20) AIDs, viral Fusion inhibitor LEXIVA ® GSK/Vertex HIV infection (orFosamprenavir calcium) AIDs, viral protease inhibitor Selzentry PfizerHIV infection Maraviroc; (UK 427857) AIDs, (CCR5 antagonist, indevelopment) Trizivir ® GSK HIV infection AIDs, (three drug combination)Sch-417690 (vicriviroc) Schering-Plough HIV infection AIDs, (CCR5antagonist, in development) TAK-652 Takeda HIV infection AIDs, (CCR5antagonist, in development) GSK 873140 GSK/ONO HIV infection (ONO-4128)AIDs, (CCR5 antagonist, in development) Integrase Inhibitor Merck HIVinfection MK-0518 AIDs Raltegravir TRUVADA ® Gilead Combination ofTenofovir disoproxil fumarate salt (VIREAD ®) and EMTRIVA ®(Emtricitabine) Integrase Inhibitor Gilead/Japan Tobacco HIV InfectionGS917/JTK-303 AIDs Elvitegravir in development Triple drug combinationGilead/Bristol-Myers Squibb Combination of Tenofovir ATRIPLA ®disoproxil fumarate salt (VIREAD ®), EMTRIVA ® (Emtricitabine), andSUSTIVA ® (Efavirenz) FESTINAVIR ® Oncolys BioPharma HIV infection4′-ethynyl-d4T BMS AIDs in development CMX-157 Chimerix HIV infectionLipid conjugate of AIDs nucleotide tenofovir GSK1349572 GSK HIVinfection Integrase inhibitor AIDs dolutegravir S/GSK1265744 GSK HIVinfection Integrase inhibitor AIDs

IMMUNOMODULATORS Drug Name Manufacturer Indication AS-101 Wyeth-AyerstAIDS Bropirimine Pharmacia Upjohn Advanced AIDS Acemannan CarringtonLabs, Inc. AIDS, ARC (Irving, TX) CL246,738 Wyeth AIDS, Kaposi's LederleLabs sarcoma FP-21399 Fuki ImmunoPharm Blocks HIV fusion with CD4+ cellsGamma Interferon Genentech ARC, in combination w/TNF (tumor necrosisfactor) Granulocyte Genetics Institute AIDS Macrophage Colony SandozStimulating Factor Granulocyte Hoechst-Roussel AIDS Macrophage ColonyImmunex Stimulating Factor Granulocyte Schering-Plough AIDS, MacrophageColony combination Stimulating Factor w/AZT HIV Core Particle RorerSeropositive HIV Immunostimulant IL-2 Cetus AIDS, in combinationInterleukin-2 w/AZT IL-2 Hoffman-LaRoche AIDS, ARC, HIV, inInterleukin-2 Immunex combination w/AZT IL-2 Chiron AIDS, increase inInterleukin-2 CD4 cell counts (aldeslukin) Immune Globulin CutterBiological Pediatric AIDS, in Intravenous (Berkeley, CA) combinationw/AZT (human) IMREG-1 Imreg AIDS, Kaposi's (New Orleans, LA) sarcoma,ARC, PGL IMREG-2 Imreg AIDS, Kaposi's (New Orleans, LA) sarcoma, ARC,PGL Imuthiol Diethyl Merieux Institute AIDS, ARC Dithio CarbamateAlpha-2 Schering Plough Kaposi's sarcoma Interferon w/AZT, AIDSMethionine- TNI Pharmaceutical AIDS, ARC Enkephalin (Chicago, IL) MTP-PECiba-Geigy Corp. Kaposi's sarcoma Muramyl-Tripeptide Granulocyte AmgenAIDS, in combination Colony Stimulating w/AZT Factor Remune ImmuneResponse Immunotherapeutic Corp. rCD4 Genentech AIDS, ARC RecombinantSoluble Human CD4 rCD4-IgG AIDS, ARC hybrids Recombinant Biogen AIDS,ARC Soluble Human CD4 Interferon Hoffman-La Roche Kaposi's sarcoma Alfa2a AIDS, ARC, in combination w/AZT SK&F106528 Smith Kline HIV infectionSoluble T4 Thymopentin Immunobiology HIV infection Research Institute(Annandale, NJ) Tumor Necrosis Genentech ARC, in combination Factor; TNFw/gamma Interferon

ANTI-INFECTIVES Drug Name Manufacturer Indication Clindamycin withPharmacia Upjohn PCP Primaquine Fluconazole Pfizer Cryptococcalmeningitis, candidiasis Pastille Squibb Corp. Prevention of NystatinPastille oral candidiasis Ornidyl Merrell Dow PCP EflornithinePentamidine LyphoMed PCP treatment Isethionate (IM & IV) (Rosemont, IL)Trimethoprim Antibacterial Trimethoprim/sulfa Antibacterial PiritreximBurroughs Wellcome PCP treatment Pentamidine Fisons Corporation PCPprophylaxis Isethionate for Inhalation Spiramycin Rhone-PoulencCryptosporidial diarrhea Intraconazole- Janssen-Pharm. Histoplasmosis;R51211 cryptococcal meningitis Trimetrexate Warner-Lambert PCPDaunorubicin NeXstar, Sequus Kaposi's sarcoma Recombinant Human OrthoPharm. Corp. Severe anemia Erythropoietin assoc. with AZT therapyRecombinant Human Serono AIDS-related Growth Hormone wasting, cachexiaMegestrol Acetate Bristol-Myers Squibb Treatment of anorexia assoc.W/AIDS Testosterone Alza, Smith Kline AIDS-related wasting Total EnteralNorwich Eaton Diarrhea and Nutrition Pharmaceuticals malabsorptionrelated to AIDS

Additionally, the compounds of the disclosure herein set forth may beused in combination with HIV entry inhibitors. Examples of such HIVentry inhibitors are discussed in DRUGS OF THE FUTURE 1999, 24(12), pp.1355-1362; CELL, Vol. 9, pp. 243-246, Oct. 29, 1999; and DRUG DISCOVERYTODAY, Vol. 5, No. 5, May 2000, pp. 183-194 and Inhibitors of the entryof HIV into host cells. Meanwell, Nicholas A.; Kadow, John F., CurrentOpinion in Drug Discovery & Development (2003), 6(4), 451-461.Specifically the compounds can be utilized in combination withattachment inhibitors, fusion inhibitors, and chemokine receptorantagonists aimed at either the CCR5 or CXCR4 coreceptor. HIV attachmentinhibitors are also set forth in U.S. Pat. Nos. 7,354,924 and 7,745,625.

It will be understood that the scope of combinations of the compounds ofthis application with AIDS antivirals, immunomodulators,anti-infectives, HIV entry inhibitors or vaccines is not limited to thelist in the above Table but includes, in principle, any combination withany pharmaceutical composition useful for the treatment of AIDS.

Preferred combinations are simultaneous or alternating treatments with acompound of the present disclosure and an inhibitor of HIV proteaseand/or a non-nucleoside inhibitor of HIV reverse transcriptase. Anoptional fourth component in the combination is a nucleoside inhibitorof HIV reverse transcriptase, such as AZT, 3TC, ddC or ddl. A preferredinhibitor of HIV protease is REYATAZ® (active ingredient Atazanavir).Typically a dose of 300 to 600 mg is administered once a day. This maybe co-administered with a low dose of Ritonavir (50 to 500 mgs). Anotherpreferred inhibitor of HIV protease is KALETRA®. Another usefulinhibitor of HIV protease is indinavir, which is the sulfate salt ofN-(2(R)-hydroxy-1-(S)-indanyl)-2(R)-phenylmethyl-4-(S)-hydroxy-5-(1-(4-(3-pyridyl-methyl)-2(S)—N′-(t-butylcarboxamido)-piperazinyl))-pentaneamideethanolate, and is synthesized according to U.S. Pat. No. 5,413,999.Indinavir is generally administered at a dosage of 800 mg three times aday. Other preferred protease inhibitors are nelfinavir and ritonavir.Another preferred inhibitor of HIV protease is saquinavir which isadministered in a dosage of 600 or 1200 mg tid. Preferred non-nucleosideinhibitors of HIV reverse transcriptase include efavirenz. Thesecombinations may have unexpected effects on limiting the spread anddegree of infection of HIV. Preferred combinations include those withthe following (1) indinavir with efavirenz, and, optionally, AZT and/or3TC and/or ddl and/or ddC; (2) indinavir, and any of AZT and/or ddland/or ddC and/or 3TC, in particular, indinavir and AZT and 3TC; (3)stavudine and 3TC and/or zidovudine; (4) tenofovir disoproxil fumaratesalt and emtricitabine.

In such combinations the compound(s) of the present invention and otheractive agents may be administered separately or in conjunction. Inaddition, the administration of one element may be prior to, concurrentto, or subsequent to the administration of other agent(s).

General Chemistry (Methods of Synthesis)

The present invention comprises compounds of Formula I, theirpharmaceutical formulations, and their use in patients suffering from orsusceptible to HIV infection. The compounds of Formula I also includepharmaceutically acceptable salts thereof. Procedures to constructcompounds of Formula I and intermediates useful for their synthesis aredescribed after the Abbreviations.

Abbreviations

One or more of the following abbreviations, most of which areconventional abbreviations well known to those skilled in the art, maybe used throughout the description of the disclosure and the examples:

-   RT=room temperature-   BHT=2,6-di-tert-butyl-4-hydroxytoluene-   CSA=camphorsulfonic acid-   LDA=lithium diisopropylamide-   KHMDS=potassium bis(trimethylsilyl)amide-   SFC=supercritical fluid chromatography-   Quant=quantitative-   TBDMS=tert-butyldimethylsilane-   PTFE=polytetrafluoroethylene-   NMO=4-methylmorpholine-N-oxide-   THF=tetrahydrofuran-   TLC=thin layer chromatography-   DCM=dichloromethane-   DCE=dichloroethane-   TFA=trifluoroacetic acid-   LCMS=liquid chromatography mass spectroscopy-   Prep=preparative-   HPLC=high performance liquid chromatography-   DAST=(diethylamino)sulfur trifluoride-   TEA=triethylamine-   DIPEA=N,N-diisopropylethylamine-   HATU=[O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium    hexafluorophosphate]-   DCC=N,N′-dicyclohexylcarbodiimide-   DMAP=dimethylaminopyridine-   TMS=trimethylsilyl-   NMR=nuclear magnetic resonance-   DPPA=diphenyl phosphoryl azide-   AIBN=azobisisobutyronitrile-   TBAF=tetrabutylaminonium fluoride-   DMF=dimethylformamide-   TBTU=O-(benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium    tetrafluoroborate-   Min(s)=minute(s)-   h=hour(s)-   sat.=saturated-   TEA=triethylamine-   EtOAc=ethyl acetate-   TFA=trifluoroacetic acid-   PCC=pyridinium chlorochromate-   TLC=thin layer chromatography-   Tf₂NPh=(trifluoromethylsulfonyl)methanesulfonamide-   dioxane=1,4-dioxane-   PG=protective group-   atm=atmosphere(s)-   mol=mole(s)-   mmol=milimole(s)-   mg=milligram(s)-   μg=microgram(s)-   μl=microliter(s)-   μm=micrometer(s)-   mm=millimeter(s)-   Rpm=revolutions per minute-   SM=starting material-   TLC=thin layer chromatography-   AP=area percentage-   Equiv.=equivalent(s)-   DMP=Dess-Martin periodinane-   TMSCl=trimethylsilyl chloride-   TBSCl=tert-Butyldimethylsilyl chloride-   TBSOTf=trimethylsilyl trifluoromethanesulfonate-   PhMe=toluene-   PhNTf₂=N-Phenyl-bis(trifluoromethanesulfonimide)-   S-Phos=2-Dicyclohexylphosphino-2′,6′-dimethoxybiphenyl-   TFDO=methyl(trifluoromethyl)dioxirane-   TEMPO=2,2,6,6-tetramethylpiperidinyloxy-   DI=deionized water

The terms “C-3” and “C-28” refer to certain positions of a triterpenecore as numbered in accordance with IUPAC rules (positions depictedbelow with respect to an illustrative triterpene: betulin):

The same numbering is maintained when referring to the compound seriesin schemes and general descriptions of methods.

EXAMPLES

The following examples illustrate typical syntheses of the compounds ofFormula I as described generally above. These examples are illustrativeonly and are not intended to limit the disclosure in any way. Thereagents and starting materials are readily available to one of ordinaryskill in the art.

Chemistry

Typical Procedures and Characterization of Selected Examples:

Unless otherwise stated, solvents and reagents were used directly asobtained from commercial sources, and reactions were performed under anitrogen atmosphere. Flash chromatography was conducted on Silica gel 60(0.040-0.063 particle size; EM Science supply). ¹H NMR spectra wererecorded on Bruker DRX-500f at 500 MHz (or Bruker AV 400 MHz, BrukerDPX-300B, or Varian Gemini 300 at 300 MHz as stated). The chemicalshifts were reported in ppm on the δ scale relative to δTMS=0. Thefollowing internal references were used for the residual protons in thefollowing solvents: CDCl₃ (δ_(H) 7.26), CD₃OD (δ_(H) 3.30), acetic-d4(Acetic Acid d₄) (δ_(H) 11.6, 2.07), DMSO mix or DMSO-D6-CDCl₃ (δ_(H)2.50 and 8.25) (ratio 75%:25%), and DMSO-D6 (δ_(H) 2.50). Standardacronyms were employed to describe the multiplicity patterns: s(singlet), br. s (broad singlet), d (doublet), t (triplet), q (quartet),m (multiplet), b (broad), app (apparent). The coupling constant (J) isin Hertz. All Liquid Chromatography (LC) data were recorded on aShimadzu LC-10AS liquid chromatograph using a SPD-10AV UV-Vis detectorwith Mass Spectrometry (MS) data determined using a Micromass Platformfor LC in electrospray mode.

LCMS Methods

LCMS Method 1:

Start % B=0

Final % B=100

Gradient Time=2 min

Flow Rate=1 mL/min

Wavelength=220 nm

Solvent A=10% MeOH-90% H₂O-0.1% TFA

Solvent B=90% MeOH-10% H₂O-0.1% TFA

Column=Phenomenex C18 2.0×30 mm 3 μm

LCMS Method 2:

Start % B=20

Final % B=100

Gradient Time=3 min

Flow Rate=0.6 mL/min

Wavelength=220 nm

Solvent A=10% MeOH-90% H₂O-0.1% TFA

Solvent B=90% MeOH-10% H₂O-0.1% TFA

Column=Xbridge Phenyl 2.1×50 mm 2.5 μm

LCMS Method 3:

Start % B=20

Final % B=100

Gradient Time=2 min

Flow Rate=0.6 mL/min

Wavelength=220 nm

Solvent A=10% MeOH-90% H₂O-0.1% TFA

Solvent B=90% MeOH-10% H₂O-0.1% TFA

Column=Xbridge Phenyl 2.1×50 mm 2.5 μm

LCMS Method 4:

Start % B=0

Final % B=100

Gradient Time=4 min

Flow Rate=0.8 mL/min

Wavelength=220 nm

Solvent A=10% MeOH-90% H₂O-0.1% TFA

Solvent B=90% MeOH-10% H₂O-0.1% TFA

Column=Phenomenex C18 2.0×50 mm 3 μm

LCMS Method 5:

Start % B=20

Final % B=100

Gradient Time=3 min

Flow Rate=0.8 mL/min

Wavelength=220 nm

Solvent A=10% MeOH-90% H₂O-0.1% TFA

Solvent B=90% MeOH-10% H₂O-0.1% TFA

Column=Phenomenex C18 2.0×50 mm 3 μm

LCMS Method 6:

Start % B=20

Final % B=100

Gradient Time=2 min

Flow Rate=0.8 mL/min

Wavelength=220 nm

Solvent A=10% MeOH-90% H2O-0.1% TFA

Solvent B=90% MeOH-10% H2O-0.1% TFA

Column=Phenomenex C18 2.0×50 mm 3 μm

LCMS Method 7:

Start % B=20

Final % B=100

Gradient Time=2 min

Flow Rate=0.5 mL/min

Wavelength=220 nm

Solvent A=10% MeOH-90% H2O-0.1% TFA

Solvent B=90% MeOH-10% H2O-0.1% TFA

Column=Xbridge Phenyl 2.1×50 mm 2.5 μm

LCMS Method 8:

Start % B=20

Final % B=100

Gradient Time=2 min

Flow Rate=0.8 mL/min

Wavelength=220 nm

Solvent A=10% MeOH-90% H2O-0.1% TFA

Solvent B=90% MeOH-10% H2O-0.1% TFA

Column=Xbridge Phenyl 2.1×50 mm 2.5 μm

LCMS Method 9:

Start % B=0

Final % B=100

Gradient Time=2 min

Flow Rate=1.0 mL/min

Wavelength=220 nm

Solvent A=5% MeCN-95% H2O-10 mM Ammonium Acetate

Solvent B=95% MeCN-5% H2O-10 mM Ammonium Acetate

Column=PHENOMENEX-LUNA C18 2.0×30 mm 3 μm

LCMS Method 10:

Start % B=0

Final % B=100

Gradient Time=4 min

Flow Rate=0.6 mL/min

Wavelength=220 nm

Solvent A=10% MeOH-90% H2O-0.1% TFA

Solvent B=90% MeOH-10% H2O-0.1% TFA

Column=Xbridge Phenyl 2.1×50 mm 2.5 μm

LCMS Method 11:

Start % B=0

Final % B=100

Gradient Time=4 min

Flow Rate=0.8 mL/min

Wavelength=220 nm

Solvent A=10% MeOH-90% H2O-0.1% TFA

Solvent B=90% MeOH-10% H2O-0.1% TFA

Column=Phenomenex C18 2.0×50 mm 3 μm

LCMS Method 12:

Start % B=40

Final % B=60

Gradient Time=4 min

Flow Rate=0.8 mL/min

Wavelength=254 nm

Solvent A=10% MeOH-90% H2O-0.1% TFA

Solvent B=90% MeOH-10% H2O-0.1% TFA

Column=Xbridge Phenyl 2.1×50 mm 2.5 μm

LCMS Method 13:

Start % B=35

Final % B=100

Gradient Time=4 min

Flow Rate=0.8 mL/min

Wavelength=220 nm

Solvent A=10% MeOH-90% H2O-0.1% TFA

Solvent B=90% MeOH-10% H2O-0.1% TFA

Column=Phenomenex C18 2.0×50 mm 3 μm

LCMS Method 14

Conditions: 0% B→100% B over 4 minute gradient; hold at 100% B for 1 min

Solvent A: 90% water, 10% methanol, 0.1% TFA

Solvent B: 10% water, 90% methanol, 0.1% TFA

Column: Phenomenex Luna C18, 3 mm, 2.0×50 mm

Flow Rate: 1 mL/min

Detector Wavelength: 220 nm

LCMS Method 15

Conditions: 0% B→100% B over 2 minute gradient; hold at 100% B for 1 min

Solvent A: 90% water, 10% methanol, 0.1% TFA

Solvent B: 10% water, 90% methanol, 0.1% TFA

Column: Phenomenex Luna C18, 2.0×50 mm, 3 μm

Flow Rate: 1 mL/min

Detector Wavelength: 220 nm

LCMS Method 16

Start % B=2, Final % B=98 over 1.5 minute gradient; hold at 98% B for0.5 min

Flow Rate=0.8 mL/min

Detector Wavelength=220 nm

Solvent A=100% water, 0.05% TFA

Solvent B=100% acetonitrile, 0.05% TFA

Column=Waters Aquity UPLC BEH C18 2.1×50 mm 1.7 um

Oven temp=40° C.

LCMS Method 17

Start % B=2, Final % B=98 over 3 minute gradient; hold at 98% B for 1min

Flow Rate=0.8 mL/min

Detector Wavelength=220 nm

Solvent A=100% water, 0.05% TFA

Solvent B=100% acetonitrile, 0.05% TFA

Column=Waters Aquity UPLC BEH C18, 2.1×50 mm, 1.7 μm

Oven temp=40° C.

LCMS Method 18

Start % B=0, Final % B=100 over 4 minute gradient; hold at 100% B for 1min

Flow Rate=0.8 mL/min

Detector Wavelength=220 nm

Solvent A=95% water, 5% acetonitrile, 10 mM aminonium acetate

Solvent B=5% water, 95% acetonitrile, 10 mM aminonium acetate

Column=Phenomenex Luna C18, 50×2 mm, 3 μm

Oven temp=40° C.

LCMS Method 19

Start % B=2, Final % B=98 over 4 minute gradient; hold at 98% B for 1min

Flow Rate=0.8 mL/min

Detector Wavelength=220 nm

Solvent A=100% water, 0.05% TFA

Solvent B=100% acetonitrile, 0.05% TFA

Column=Waters Aquity UPLC BEH C18, 2.1×50 mm, 1.7 μm

Oven temp=40° C.

LCMS Method 20

Start % B=2, Final % B=98 over 2 minute gradient; hold at 98% B for 1min

Flow Rate=0.8 mL/min

Detector Wavelength=220 nm

Solvent A=100% water, 0.05% TFA

Solvent B=100% acetonitrile, 0.05% TFA

Column=Waters Aquity UPLC BEH C18, 2.1×50 mm, 1.7 μm

Oven temp=40° C.

LCMS Method 21

Start % B=0, Final % B=100 over 2 minute gradient; hold at 100% B for 3min

Flow Rate=0.8 mL/min

Detector Wavelength=220 nm

Solvent A=95% water, 5% acetonitrile, 10 mM aminonium acetate

Solvent B=5% water, 95% acetonitrile, 10 mM aminonium acetate

Column=Phenomenex Luna C18, 50×2 mm, 3 μm

Oven temp=40° C.

Preparative HPLC Methods

Preparative HPLC Method 1

Conditions: 30% B→100% B over 20 minute gradient; hold at 100% B for 4min

Solvent A: 5% acetonitrile, 95% water, 0.1% TFA

Solvent B: 95% acetonitrile, 5% water 0.1% TFA

Column: Waters Xbridge 30×100 mm, 5 μm

Flow Rate: 40 mL/min

Detector Wavelength: 220 nm

Preparative HPLC Method 2

Conditions: 10% B→100% B over 25 minute gradient

Solvent A: 5% acetonitrile, 95% water, 0.1% TFA

Solvent B: 95% acetonitrile, 5% water 0.1% TFA

Column: Waters Sunfire 30×150 mm, 5 um

Flow Rate: 40 mL/min

Detector Wavelength: 220 nm

Preparative HPLC Method 3

Conditions: 10% B→100% B over 20 minute gradient; hold at 100% B for 5min

Solvent A: 5% acetonitrile, 95% water, 0.1% TFA

Solvent B: 95% acetonitrile, 5% water 0.1% TFA

Column: Waters Sunfire 30×150 mm, 5 um

Flow Rate: 40 mL/min

Detector Wavelength: 220 nm

Preparative HPLC Method 4

Conditions: 30% B→100% B over 20 minute gradient; hold at 100% B for 5min

Solvent A: 5% acetonitrile, 95% water, 0.1% TFA

Solvent B: 95% acetonitrile, 5% water 0.1% TFA

Column: Waters Sunfire 30×150 mm, 5 um

Flow Rate: 40 mL/min

Detector Wavelength: 220 nm

Preparative HPLC Method 5:

Start % B=20, Final % B=100 over 10 min gradient, hold at 100% B for 4min

Flow Rate=50 ml/min

Wavelength=220

Solvent Pair=Water-acetonitrile-TFA

Solvent A=90% Water-10% acetonitrile-0.1% TFA

Solvent B=10% Water-90% acetonitrile-0.1% TFA

Column=Waters Sunfire C18, 5 μm, 30×150 mm

Preparative HPLC Method 6

Conditions: 0% B→100% B over 20 minute gradient

Solvent A: 10% acetonitrile, 90% water, 0.1% TFA

Solvent B: 90% acetonitrile, 10% water 0.1% TFA

Column: Waters Sunfire C18, 30×150 mm, 5 μm

Flow Rate: 50 mL/min

Detector Wavelength: 220 nm

Preparative HPLC Method 7

Conditions: 30% B→100% B over 20 minute gradient

Solvent A: 10% acetonitrile, 90% water, 0.1% TFA

Solvent B: 90% acetonitrile, 10% water 0.1% TFA

Column: Waters Sunfire C18, 30×150 mm, 5 μm

Flow Rate: 50 mL/min

Detector Wavelength: 220 nm

Preparative HPLC Method 8

Conditions: 20% B→100% B over 15 minute gradient

Solvent A: 10% acetonitrile, 90% water, 0.1% TFA

Solvent B: 90% acetonitrile, 10% water 0.1% TFA

Column: Waters Sunfire C18, 30×150 mm, 5 μm

Flow Rate: 50 mL/min

Detector Wavelength: 220 nm

Preparative MPLC Methods

Preparative MPLC Method 1

Conditions: 30% B for 1 column volume, 30% B to 80% B gradient over 7column volumes, 80% B to 100% B gradient over 0.5 column volumes, 100% Bfor 2 column volumes

Solvent A=5% acetonitrile, 95% water, 0.1% TFA

Solvent B=95% acetonitrile, 5% water 0.1% TFA

Column=Redi Sep Gold (150 g)

Flow Rate=60 mL/min

Detector Wavelength=220 nm

Preparative MPLC Method 2

Conditions: 30% B for 1 column volume, 30% B to 80% B gradient over 10column

volumes, 100% B for 2 column volumes

Solvent A=5% acetonitrile, 95% water, 0.1% TFA

Solvent B=95% acetonitrile, 5% water 0.1% TFA

Column=Redi Sep Gold (150 g)

Flow Rate=60 mL/min

Detector Wavelength=220 nm

Analytical HPLC Methods

Analytical HPLC Method 1

Conditions: 10% B→100% B over 15 min gradient; hold at 100% B for 10 min

Solvent A: 10% methanol, 90% water, 0.1% TFA

Solvent B: 90% methanol, 10% water, 0.1% TFA

Column: Waters Sunfire C18, 4.6×150 mm, 3.5 mm

Flow Rate: 1 mL/min

Detector Wavelength: 220 nm

Analytical HPLC Method 2

Conditions: 10% B→100% B over 15 min gradient; hold at 100% B for 10 min

Solvent A: 10% methanol, 90% water, 0.1% TFA

Solvent B: 90% methanol, 10% water, 0.1% TFA

Column: Waters Xbridge phenyl, 4.6×150 mm, 3.5 mm

Flow Rate: 1 mL/min

Detector Wavelength: 220 nm

Analytical HPLC Method 3

Conditions: 10% B→100% B over 15 min gradient; hold at 100% B for 10 min

Solvent A: 5% acetonitrile, 95% water, 0.1% TFA

Solvent B: 95% acetonitrile, 5% water, 0.1% TFA

Column: Waters Sunfire C18, 3.0×150 mm, 3.5 um

Flow Rate: 0.5 mL/min

Detector Wavelength: 220 nm

Analytical HPLC Method 4

Conditions: 10% B→100% B over 15 min gradient; hold at 100% B for 10 min

Solvent A: 5% acetonitrile, 95% water, 0.1% TFA

Solvent B: 95% acetonitrile, 5% water, 0.1% TFA

Column: Waters Xbridge phenyl, 3.0×150 mm, 3.5 um

Flow Rate: 0.5 mL/min

Detector Wavelength: 220 nm

Preparation of Intermediates

Intermediate 1. Preparation of ethyl1,4-dioxaspiro[4.5]decane-8-carboxylate

A mixture of ethyl 4-oxocyclohexanecarboxylate (12.7 g, 75 mmol),ethylene glycol (21 ml, 373 mmol), (1S)-(+)-10-camphorsulfonic acid(0.175 g, 0.75 mmol) and anhydrous toluene (300 mL) was refluxed with aDean-Stark water trap for 8 hours. The mixture was quenched with 100 mLsaturated sodium bicarbonate solution and was vigorously stirred. Theseparated organic phase was washed with water (100 mL), dried overNa₂SO₄ and concentrated in vacuo. The crude product was purified bysilica gel column eluted with 0-15% ethyl acetate/hexanes to give thedesired product as an oil (15.9 g, 99%). ¹H NMR (400 MHz, CHLOROFORM-d)δ 4.13 (q, J=7.2 Hz, 2H), 3.95 (s, 4H), 2.34 (tt, J=10.4, 4.0 Hz, 1H),1.98-1.90 (m, 2H), 1.87-1.75 (m, 4H), 1.61-1.51 (m, 2H), 1.25 (t, J=7.2Hz, 3H).

Intermediate 2. Preparation of ethyl8-formyl-1,4-dioxaspiro[4.5]decane-8-carboxylate

To a solution of ethyl 1,4-dioxaspiro[4.5]decane-8-carboxylate (21 g, 98mmol) in THF (150 mL) at −78° C. was added 2M LDA (64 mL, 127 mmol)dropwise. The resulting solution was stirred at −78° C. for 1 h, then inan ice bath for 1.5 h. The reaction mixture was chilled back to −78° C.and molecular sieves were added. Dried ethyl formate (12 mL, 147 mmol)was added dropwise slowly over 1 h. The reaction mixture was stirred at−78° C. for 1 h. The cold bath was removed and the reaction was quenchedwith a saturated solution of NH₄Cl in 0.5 N HCl (250 mL) dropwise. Themixture was extracted with EtOAc (3×200 mL). The combined organic layerwas washed with saturated solution of NH₄Cl in 0.5 N HCl (200 mL), brine(200 mL), dried over Na₂SO₄, and concentrated in vacuo. The crudeproduct was purified by silica gel column eluted with 0-20% ethylacetate/hexanes to give the desired product as an oil (9.3 g, 39%). ¹HNMR (400 MHz, CHLOROFORM-d) δ 9.54 (s, 1H), 4.21 (q, J=7.1 Hz, 2H),3.98-3.90 (m, 4H), 2.25-2.16 (m, 2H), 2.10-2.01 (m, 2H), 1.74-1.60 (m,4H), 1.27 (t, J=7.2 Hz, 3H).

Intermediate 3. Preparation of ethyl8-(hydroxymethyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate

To a solution of the ethyl8-formyl-1,4-dioxaspiro[4.5]decane-8-carboxylate (1.0 g, 4.13 mmol) inEtOH (10 mL) at 0° C. was added NaBH₄ (0.187 g, 4.95 mmol). The mixturewas stirred at 0° C. for 1 h. The reaction was quenched with saturatedNH₄Cl (10 mL) and was then diluted with H₂O until dissolved. The mixturewas extracted with EtOAc (3×50 mL), washed with brine (50 mL), driedover Na₂SO₄ and concentrated in vacuo. The crude product was purified bysilica gel column eluted with 0-25% ethyl acetate/hexanes to give thedesired product as an oil (0.86 g, 85%). ¹H NMR (400 MHz, CHLOROFORM-d)δ 4.21 (q, J=7.1 Hz, 2H), 3.99-3.91 (m, 4H), 3.65 (d, J=6.5 Hz, 2H),2.19-2.11 (m, 2H), 1.68 (dd, J=6.8, 5.5 Hz, 4H), 1.63-1.57 (m, 2H), 1.29(t, J=7.0 Hz, 3H).

Intermediate 4. Preparation of ethyl8-((benzoyloxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate

To a solution of ethyl8-(hydroxymethyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate (3.0 g, 12.3mmol) in pyridine (60 mL) was added DMAP (0.3 g, 2.5 mmol). The mixturewas heated to 50° C. and benzoic anhydride (3.1 g, 13.5 mmol) was added.The reaction mixture was stirred at 50° C. for 3 h. The reaction mixturewas concentrated in vacuo. The residue was dissolved in EtOAc (50 mL),washed with brine (50 mL), dried over Na₂SO₄, and concentrated in vacuo.The crude product was purified by silica gel column eluted with 0-20%hexane/EtOAc to give the desired product as an oil (4.3 g, 100%). ¹H NMR(400 MHz, CHLOROFORM-d) δ 8.01 (dd, J=8.4, 1.4 Hz, 2H), 7.60-7.54 (m,1H), 7.47-7.40 (m, 2H), 4.35 (s, 2H), 4.20 (q, J=7.2 Hz, 2H), 3.99-3.92(m, 4H), 2.36-2.23 (m, 2H), 1.76-1.63 (m, 6H), 1.24 (t, J=12 Hz, 3H).

Intermediate 5. Preparation of(1-(ethoxycarbonyl)-4-oxocyclohexyl)methyl benzoate

A solution of ethyl8-((benzoyloxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate (4.3 g,12.4 mmol) in acetone (120 mL) and 0.5N HCl (24.8 mL, 12.4 mmol) wasstirred at 50° C. overnight. The reaction mixture was neutralized withsaturated aqueous Na₂CO₃ and partially concentrated in vacuo to removeacetone. The residue was diluted with H₂O (50 mL) and extracted withEtOAc (3×100 mL). The combined organic layers were washed brine (50 mL),dried over Na₂SO₄, and concentrated in vacuo. The crude product waspurified by silica gel column eluted with 0-30% hexane/EtOAc to give thedesired product as an oil (3.8 g, 100%). ¹H NMR (400 MHz, CHLOROFORM-d)δ 8.01 (d, J=7.6 Hz, 2H), 7.62-7.55 (m, 1H), 7.49-7.42 (m, 2H), 4.45 (s,2H), 4.28 (q, J=7.1 Hz, 2H), 2.61-2.48 (m, 4H), 2.47-2.37 (m, 2H),1.91-1.79 (m, 2H), 1.28 (t, J=1.1 Hz, 3H).

Intermediate 6. Preparation of(1-(ethoxycarbonyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-en-1-yl)methylbenzoate

A solution of (1-(ethoxycarbonyl)-4-oxocyclohexyl)methyl benzoate (3.8g, 12.4 mmol) and1,1,1-trifluoro-N-phenyl-N-((trifluoromethyl)sulfonyl)methanesulfonamide(4.95 g, 13.8 mmol) in THF (120 mL) was cooled to −78° C. To thissolution was added KHMDS (1 M in THF) (16.4 mL, 16.4 mmol). Theresulting solution was stirred at −78° C. for 2 h. The reaction wasquenched with saturated aqueous NH₄Cl (50 mL), extracted with EtOAc(3×100 mL). The combined organic layers were washed with brine (50 mL),dried over Na₂SO₄, and concentrated in vacuo. The crude product waspurified by silica gel column eluted with 0-20% ethyl acetate/hexanes togive the desired product (3.8 g, 69%) as an oil. ¹H NMR (400 MHz,CHLOROFORM-d) δ 8.00 (dd, J=8.4, 1.1 Hz, 2H), 7.62-7.56 (m, 1H),7.49-7.44 (m, 2H), 5.80 (td, J=3.2, 1.6 Hz, 1H), 4.46-4.40 (m, 2H), 4.21(qd, J=7.1, 2.1 Hz, 2H), 2.93-2.83 (m, 1H), 2.59-2.27 (m, 4H), 1.99-1.90(m, 1H), 1.25 (t, J=7.2 Hz, 3H).

Intermediate 7.(1-(ethoxycarbonyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-en-1-yl)methylbenzoate

A mixture of(1-(ethoxycarbonyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-en-1-yl)methylbenzoate (3.8 g, 8.7 mmol), bis(pinacolato)diboron (2.4 g, 9.5 mmol),potassium acetate (2.6 g, 26.0 mmol) and PdCl₂(dppf)-CH₂Cl₂ adduct (0.2g, 0.260 mmol) in 1,4-dioxane (80 mL) was cooled to −78° C. Three cyclesof evacuating the flask and purging with nitrogen were performed. Themixture was stirred at 70° C. for 3 h. The mixture was diluted withwater (50 mL) and extracted with ethyl acetate (3×100 mL). The combinedorganic layers were washed with brine (50 mL), dried over Na₂SO₄, andconcentrated in vacuo. The crude product was purified by silica gelcolumn eluted with 0-20% ethyl acetate/hexanes to give the desiredproduct (5.8 g, 67%) as an oil. ¹H NMR (400 MHz, CHLOROFORM-d) δ 8.00(dd, J=8.4, 1.4 Hz, 2H), 7.59-7.54 (m, 1H), 7.46-7.41 (m, 2H), 6.54 (dt,J=3.6, 1.9 Hz, 1H), 4.44 (d, J=10.8 Hz, 1H), 4.39 (d, J=10.8 Hz, 2H),4.17 (q, J=7.2 Hz, 2H), 2.77-2.68 (m, 1H), 2.29-2.20 (m, 3H), 2.05-1.97(m, 1H), 1.92-1.83 (m, 1H), 1.27 (s, 12H), 1.22 (t, J=7.2 Hz, 3H).

Intermediates 8 and 9. Chiral separation of(S)-(1-(ethoxycarbonyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-en-1-yl)methylbenzoate and (R)-(1-(ethoxycarbonyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-en-1-yl)methylbenzoate

The racemic mixture was separated by supercritical fluid chromatography(SFC) to give(S)-(1-(ethoxycarbonyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-en-1-yl)methylbenzoate and(R)-(1-(ethoxycarbonyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-en-1-yl)methylbenzoate.

SFC Experimental Details: Column: ChiralCel OJ-H, 30 × 250 mm, 5 μmMobile Phase: 5% MeOH/95% CO2 Pressure: 100 bar Temperature: 40° C. FlowRate: 70 mL/min UV: 225 nm Injection: 0.50 mL (~100 mg/mL inIPA:ACN:MeOH, 2:2:1) Fraction Collection: Slope & Level (w/6 mL/min MeOHmake-up): Peak 1 window: 3.00′-4.50′ Peak 2 window: 3.80′-7.00′

Intermediate 10. Preparation of ethyl8-(((methylsulfonyl)oxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate

To vacuum dried ethyl8-(hydroxymethyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate (280 mg, 1.146mmol) in DCM (2 mL) was added N,N-diisopropylethylamine (0.299 mL, 1.719mmol) under nitrogen. The clear solution was chilled in an ice bathuntil cold. To this was added, dropwise, neat methanesulfonyl chloride(0.106 mL, 1.375 mmol) and the resulting solution was stirred in the icebath and allowed to reach RT overnight. The crude reaction mixture waspurified on silica gel column eluted with 50% ethyl acetate/hexanes togive the desired product (304 mg, 82%). NMR (400 MHz, CHLOROFORM-d) δ4.26-4.17 (m, 4H), 3.97-3.93 (m, 4H), 3.00 (s, 3H), 2.24-2.15 (m, 2H),1.73-1.61 (m, 6H), 1.29 (t, J=7.2 Hz, 3H).

General Procedure A: Preparation of C-3 α-SubstitutedCyclohexenecarboxylic Acid Derivatives.

Step 1: Preparation of Ether.

Step 1-A: To a solution of ethyl8-(hydroxymethyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate (intermediate3) (1 eq), reactant Ar—OH (1 eq) and triphenylphosphine (1.2 eq) in THFwas added diisopropyl diazene-1,2-dicarboxylate (1.2 eq) dropwise undernitrogen. The resulting solution was stirred at RT for 1 h, then at 50°C. for 3 days. The reaction mixture was diluted with saturated NH₄Cl,extracted with EtOAc, washed with brine, dried over Na₂SO₄, andconcentrated in vacuo. The crude product was purified by silica gelcolumn eluted with ethyl acetate/hexanes to give the desired product.

Step 1-B: A mixture of ethyl8-(((methylsulfonyl)oxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate(1 eq), cesium carbonate (2.15 eq) and Ar—OH (3.5 eq) in acetonitrilewas stirred at 85° C. over 48 hours. The inorganic salts were removed byfiltration, and the filtrate was washed with water, extracted with ethylacetate. The combine organic phase was concentrated in vacuo. The crudeproduct was purified by silica gel column eluted with Ethylacetate/hexanes to give the desired product.

Step 2: Preparation of Ketone.

A solution of the product from step 1 (1 eq) and 0.5 N HCl (1 eq) inacetone was stirred at 50° C. for 1-2 days. The reaction mixture wasneutralized with saturated aqueous. Na₂CO₃ and partially concentrated invacuo to remove acetone. The residue was diluted with H₂O, extractedwith EtOAc, washed with brine, dried over Na₂SO₄, and concentrated invacuo. The crude product was purified by silica gel column eluted withethyl acetate/hexanes to give the desired ketone.

Step 3: Preparation of Triflate.

To a solution of ketone from step 2 (1 eq) and1,1,1-trifluoro-N-phenyl-N-((trifluoromethyl)sulfonyl)-methanesulfonamide(1.1 eq) in THF at −78° C. was added KHMDS (1 M in THF) (1.3 eq). Theresulting yellow to orange solution was stirred at −78° C. for 2 h. Thereaction was quenched with saturated aqueous NH₄Cl. The mixture wasextracted with EtOAc, washed with brine, dried over Na₂SO₄, andconcentrated in vacuo. The crude product was purified by silica gelcolumn eluted with ethyl acetate/hexanes to give the desired triflate.

Step 4: Preparation of Boronate.

In a pressure vessel, a mixture of triflate from step 3 (1 eq),bis(pinacolato)diboron (1.1 eq), KOAc (2.5 eq) and PdCl₂(dppf)-CH₂Cl₂adduct (0.03 eq) in 1,4-dioxane was flushed with nitrogen, sealed andheated at 70° C. for 2 h. The mixture was diluted with water andextracted with EtOAc. The combined organic layers were washed withbrine, dried over Na₂SO₄, and concentrated in vacuo. The crude productwas purified by silica gel column eluted with ethyl acetate/hexanes togive the desired boronate.

Step 5: Preparation of C-3 α-Substituted Cyclohexenecarboxylic Ester.

A mixture of C3-triflate (1 eq), boronate from step 4 (1 eq), Na₂CO₃H₂O(3 eq) and Pd(Ph₃P)₄ (0.06 eq) in dioxane and H₂O (4:1), was flushedwith nitrogen, sealed and heated at 70° C. for 2 h. The reaction mixturewas concentrated in vacuo, and the residue was partitioned between EtOAcand H₂O. The separated aqueous layer was extracted with EtOAc. Thecombined organic layers were washed with brine dried over Na₂SO₄, andconcentrated in vacuo. The crude product was purified by silica gelcolumn eluted with ethyl acetate/hexanes to give the desired C-3α-substituted cyclohexenecarboxylic ester.

Step 6: Preparation of Carboxylic Acid.

A solution of ester from step 5 in 1,4-dioxane, MeOH and 1N NaOH (2:1:1)was stirred at 60-70° C. for 1-2 h. The reaction mixture was purified byreverse phase preparative HPLC to give the final product.

Example 1 Preparation of4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-2-yloxy)methyl)cyclohex-3-ene-1-carboxylicacid

Step 1. Preparation of ethyl8-((pyridin-2-yloxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate

The title compound was prepared in 83% yield as an oil, following theprocedure described in general procedure A step 1-A, using pyridin-2-olas reactant. ¹H NMR (500 MHz, CHLOROFORM-d) δ 8.14 (ddd, J=5.0, 2.0, 0.8Hz, 1H), 7.59-7.53 (m, 1H), 6.87 (ddd, J=7.1, 5.1, 0.9 Hz, 1H), 6.73(dt, J=8.4, 0.8 Hz, 1H), 4.38 (s, 2H), 4.17 (q, J=7.2 Hz, 2H), 4.01-3.93(m, 4H), 2.35-2.24 (m, 2H), 1.79-1.67 (m, 6H), 1.23 (t, J=7.1 Hz, 3H).LC/MS m/z 322.10 (M+H)⁺, 1.93 min (LCMS Method 1).

Step 2. Preparation of ethyl4-oxo-1-((pyridin-2-yloxy)methyl)cyclohexane-1-carboxylate

The title compound was prepared in 99% yield as an oil, following theprocedure described in general procedure A step 2, using ethyl8-((pyridin-2-yloxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate asreactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 8.14 (ddd, J=5.1, 1.9, 0.8Hz, 1H), 7.61-7.55 (m, 1H), 6.90 (ddd, J=7.1, 5.1, 0.9 Hz, 1H), 6.74(dt, J=8.3, 0.8 Hz, 1H), 4.45 (s, 2H), 4.24 (q, J=7.0 Hz, 2H), 2.59-2.48(m, 4H), 2.46-2.37 (m, 2H), 1.94-1.83 (m, 2H), 1.26 (t, J=7.2 Hz, 3H).LC/MS m/z 278.05 (M+H)⁺, 1.74 mm (LCMS Method 1).

Step 3. Preparation of ethyl1-((pyridin-2-yloxy)methyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 110% yield (containing PhNHTf) as anoil, following the procedure described in general procedure A step 3,using ethyl 4-oxo-1-((pyridin-2-yloxy)methyl)cyclohexane-1-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 8.14 (ddd, J=5.0, 2.0, 0.8Hz, 1H), 7.62-7.55 (m, 1H), 6.90 (ddd, J=7.1, 5.1, 0.9 Hz, 1H), 6.73(dt, J=8.3, 0.8 Hz, 1H), 5.80-5.76 (m 1H), 4.45 (d, J=10.3 Hz, 1H), 4.39(d, J=10.3 Hz, 1H), 4.18 (qd, J=7.1, 1.3 Hz, 2H), 2.88-2.80 (m, 1H),2.56-2.25 (m, 4H), 2.02-1.93 (m, 1H), 1.22 (t, J=7.2 Hz, 3H). ¹⁹F NMR(376 MHz, CHLOROFORM-d) δ −73.87 (s, 3F). /LC/MS m/z 410.00 (M+H)⁺, 2.24min (LCMS Method 1).

Step 4. Preparation of ethyl1-((pyridin-2-yloxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 75% yield as an oil, following theprocedure described in general procedure A step 4, using ethyl1-((pyridin-2-yloxy)methyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-ene-1-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 8.13 (ddd, J=5.0, 2.0, 0.8Hz, 1H), 7.58-7.51 (m, 1H), 6.86 (ddd, J=7.0, 5.1, 0.9 Hz, 1H), 6.71(dt, J=8.4, 0.8 Hz, 1H), 6.57-6.53 (m, 1H), 4.42 (d, J=10.0 Hz, 1H),4.33 (d, J=10.0 Hz, 1H), 4.14 (qd, J=6.7, 1.4 Hz, 2H), 2.73 (dq, J=18.8,2.8 Hz, 1H), 2.31-2.18 (m, 3H), 2.03-1.95 (m, 1H), 1.91-1.83 (m, 1H),1.27 (s, 6H), 1.26 (s, 6H), 1.19 (t, J=7.0 Hz, 3H). LC/MS m/z 388.20(M+H)⁺, 2.22 min (LCMS Method 1).

Step 5. Preparation of ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-2-yloxy)methyl)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 71% yield as a solid, following theprocedure described in general procedure A step 5, using ethyl1-((pyridin-2-yloxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-ene-1-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 8.13 (dd, J=5.1, 1.4 Hz,1H), 7.55 (ddd, J=8.6, 7.0, 2.0 Hz, 1H), 6.85 (ddd, J=7.0, 5.3, 0.8 Hz,1H), 6.72 (d, J=8.3 Hz, 1H), 5.35 (br. s, 1H), 5.18 (d, J=5.5 Hz, 1H),4.71 (s, 1H), 4.59 (s, 1H), 4.47-4.37 (m, 2H), 4.14 ((qd, J=6.7, 1.4 Hz,2H), 3.12-2.99 (m, 8H), 2.73-2.39 (m, 6H), 2.23-0.84 (m, 27H), 1.69 (s,3H), 1.20 (t, J=7.2 Hz, 3H), 1.05 (s, 3H), 0.96-0.90 (m, 9H), 0.89 (s,3H). LC/MS m/z 830.00 (M+H)⁺, 3.74 min (LCMS Method 2).

Step 6.4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-2-yloxy)methyl)cyclohex-3-ene-1-carboxylicacid was prepared in 32% yield as a solid, following the proceduredescribed in general procedure A step 6, using ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-2-yloxy)methyl)cyclohex-3-ene-1-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 8.14 (d, J=3.8 Hz, 1H),7.59-7.52 (m, 1H), 6.89-6.84 (m, 1H), 6.73 (d, J=8.3 Hz, 1H), 5.35 (br.s, 1H), 5.21-5.16 (m, 1H), 4.71 (s, 1H), 4.60 (s, 1H), 4.50-4.38 (m,2H), 3.14-2.99 (m, 8H), 2.86-2.57 (m, 6H), 2.29-0.89 (m, 27H), 1.68 (s,3H), 1.10 (s, 3H), 0.98 (s, 3H), 0.97-0.91 (m, 6H), 0.85 (s, 3H). LC/MSm/z 802.50 (M+H)⁺, 3.56 min (LCMS Method 2).

Example 2 Preparation of4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((1-methyl-3-phenyl-1H-pyrazol-5-yl)oxy)methyl)cyclohex-3-ene-1-carboxylicacid

Step 1. Preparation of ethyl8-(((1-methyl-3-phenyl-1H-pyrazol-5-yl)oxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate

The title compound was prepared in 99% yield as an oil, following theprocedure described in general procedure A step 1-A, using1-methyl-3-phenyl-1H-pyrazol-5-ol as reactant. ¹H NMR (500 MHz,CHLOROFORM-d) δ 7.78-7.70 (m, 2H), 7.39 (t, J=7.6 Hz, 2H), 7.32-7.28 (m,1H), 5.83 (s, 1H), 4.21 (q, J=7.1 Hz, 2H), 4.12 (s, 2H), 4.01-3.94 (m,4H), 3.67 (s, 3H), 2.37-2.26 (m, 2H), 1.80-1.65 (m, 6H), 1.31-1.26 (m,3H). LC/MS m/z 401.10 (M+H)⁺, 2.17 min (LCMS Method 1).

Step 2. Preparation of ethyl1-(((1-methyl-3-phenyl-1H-pyrazol-5-yl)oxy)methyl)-4-oxocyclohexane-1-carboxylate

The title compound was prepared in 81% yield as an oil, following theprocedure described in general procedure A step 2, using ethyl8-(((1-methyl-3-phenyl-1H-pyrazol-5-yl)oxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.75-7.71 (m, 2H),7.42-7.35 (m, 2H), 7.32-7.27 (m, 1H), 5.84 (s, 1H), 4.28 (q, J=7.0 Hz,2H), 4.19 (s, 2H), 3.68 (s, 3H), 2.63-2.51 (m, 4H), 2.48-2.39 (m, 2H),1.92-1.81 (m, 2H), 1.30 (t, J=7.2 Hz, 3H). LC/MS m/z 357.15 (M+H)⁺, 1.99min (LCMS Method 1).

Step 3. Preparation of ethyl1-(((1-methyl-3-phenyl-1H-pyrazol-5-yl)oxy)methyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 68% yield as an oil, following theprocedure described in general procedure A step 3, using ethyl1-(((1-methyl-3-phenyl-1H-pyrazol-5-yl)oxy)methyl)-4-oxocyclohexane-1-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.75-7.71 (m, 2H),7.41-7.35 (m, 2H), 7.32-7.29 (m, 1H), 5.84 (s, 1H), 5.83-5.79 (m, 1H),4.25-4.10 (m, 4H), 3.67 (s, 3H), 2.92-2.82 (m, 1H), 2.59-2.25 (m, 4H),2.00 (ddd, J=13.7, 7.8, 6.4 Hz, 1H), 1.27 (t, J=7.2 Hz, 3H). ¹⁹F NMR(376 MHz, CHLOROFORM-d) 6-73.83 (s, 3F). LC/MS m/z 489.20 (M+H)⁺, 2.30min (LCMS Method 1).

Step 4. Preparation of ethyl1-(((1-methyl-3-phenyl-1H-pyrazol-5-yl)oxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 68% yield as a wax, following theprocedure described in general procedure A step 4, using ethyl1-(((1-methyl-3-phenyl-1H-pyrazol-5-yl)oxy)methyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-ene-1-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.75-7.71 (m, 2H),7.41-7.34 (m, 2H), 7.31-7.28 (m, J=7.5 Hz, 1H), 6.57-6.53 (m, 1H), 5.83(s, 1H), 4.23-4.11 (m, 4H), 3.65 (s, 3H), 2.76-2.67 (m, 1H), 2.32-2.12(m, 3H), 2.03-1.86 (m, 2H), 1.27 (s, 12H), 1.23 (t, J=7.0 Hz 3H). LC/MSm/z 467.30 (M+H)⁺, 3.58 min (LCMS Method 2).

Step 5. Preparation of ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((1-methyl-3-phenyl-1H-pyrazol-5-yl)oxy)methyl)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 59% yield as a solid, following theprocedure described in general procedure A step 5, using ethyl1-(((1-methyl-3-phenyl-1H-pyrazol-5-yl)oxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-ene-1-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.74-7.70 (m, 2H),7.40-7.33 (m, 2H), 7.31-7.25 (m, 1H), 5.83 (s, 1H), 5.36 (br. s., 1H),5.19 (d, J=4.8 Hz, 1H), 4.71 (d, J=2.0 Hz, 1H), 4.59 (s, 1H), 4.24-4.15(m, 4H), 3.65 (s, 3H), 3.10-2.98 (m, 8H), 2.74-2.43 (m, 6H), 2.32-1.02(m, 27H), 1.68 (s, 3H), 1.26 (t, J=7.0 Hz, 3H), 1.06 (s, 3H), 0.97-0.91(m, 9H), 0.86 (s, 3H). LC/MS m/z 909.60 (M+H)⁺, 3.89 min (LCMS Method2).

Step 6.4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((1-methyl-3-phenyl-1H-pyrazol-5-yl)oxy)methyl)cyclohex-3-ene-1-carboxylicacid was prepared in 81% yield as a solid, following the proceduredescribed in general procedure A step 6, using ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((1-methyl-3-phenyl-1H-pyrazol-5-yl)oxy)methyl)cyclohex-3-ene-1-carboxylateas reactant. ¹H NMR (400 MHz, METHANOL-d₄) δ 7.73-7.68 (m, 2H),7.40-7.34 (m, 2H), 7.31-7.26 (m, 1H), 6.04 (s, 1H), 5.37 (br. s., 1H),5.22 (d, J=4.5 Hz, 1H), 4.76 (s, 1H), 4.65 (s, 1H), 4.31-4.23 (m, 2H),3.64 (s, 3H), 3.20-3.04 (m, 8H), 2.92-2.61 (m, 6H), 2.24-1.10 (m, 27H),1.73 (s, 3H), 1.16 (s, 3H), 1.06 (s, 3H), 1.00 (s, 3H), 0.97 (s, 3H),0.92 (s, 3H). LC/MS m/z 881.55 (M+H)⁺, 3.77 min (LCMS Method 2).

Example 3 Preparation of4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((1-methyl-5-(trifluoromethyl)-1H-pyrazol-3-yl)oxy)methyl)cyclohex-3-ene-1-carboxylicacid

Step 1. Preparation of ethyl8-(((1-methyl-5-(trifluoromethyl)-1H-pyrazol-3-yl)oxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate

The title compound was prepared in 86% yield as an oil, following theprocedure described in general procedure A step 1-A, using1-methyl-5-(trifluoromethyl)-1H-pyrazol-3-ol as reactant. ¹H NMR (500MHz, CHLOROFORM-d) δ 5.99 (s, 1H), 4.20 (q, J=1.2 Hz, 2H), 4.19 (s, 2H),3.96 (t, J=3.0 Hz, 4H), 3.82 (s, 3H), 2.31-2.19 (m, 2H), 1.78-1.64 (m,6H), 1.26 (t, J=7.1 Hz, 3H). LC/MS m/z 393.05 (M+H)⁺, 2.18 min (LCMSMethod 1).

Step 2. Preparation of ethyl1-(((1-methyl-5-(trifluoromethyl)-1H-pyrazol-3-yl)oxy)methyl)-4-oxocyclohexane-1-carboxylate

The title compound was prepared in 98% yield as an oil, following theprocedure described in general procedure A step 2, using ethyl8-(((1-methyl-5-(trifluoromethyl)-1H-pyrazol-3-yl)oxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 5.99 (s, 1H), 4.26 (s,2H), 4.26 (q, J=7.1 Hz, 2H), 3.82 (d, J=0.8 Hz, 3H), 2.59-2.34 (m, 6H),1.92-1.79 (m, 2H), 1.28 (t, J=6.8 Hz, 3H). ¹⁹F NMR (376 MHz,CHLOROFORM-d) δ −60.88 (s, 3F). LC/MS m/z 349.15 (M+H)⁺, 2.08 min (LCMSMethod 1).

Step 3. Preparation of ethyl1-(((1-methyl-5-(trifluoromethyl)-1H-pyrazol-3-yl)oxy)methyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 70% yield as an oil, following theprocedure described in general procedure A step 3, using ethyl1-(((1-methyl-5-(trifluoromethyl)-1H-pyrazol-3-yl)oxy)methyl)-4-oxocyclohexane-1-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 5.99 (s, 1H), 5.79-5.76(m, 1H), 4.29-4.16 (m, 4H), 3.81 (d,7=0.8 Hz, 3H), 2.85-2.75 (m, 1H),2.55-2.19 (m, 4H), 2.02-1.93 (m, 1H), 1.25 (t, J=1.2 Hz, 3H). ¹⁹F NMR(376 MHz, CHLOROFORM-d) δ −60.89 (s, 3F), —73.88 (s, 3F). LC/MS m/z481.10 (M+H)⁺, 2.32 min (LCMS Method 1).

Step 4. Preparation of ethyl1-(((1-methyl-5-(trifluoromethyl)-1H-pyrazol-3-yl)oxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 79% yield as a wax, following theprocedure described in general procedure A step 4, using ethyl1-(((1-methyl-5-(trifluoromethyl)-1H-pyrazol-3-yl)oxy)methyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-ene-1-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 6.55-6.51 (m, 1H), 5.97(s, 1H), 4.25 (d, J=9.3 Hz 1H), 4.19-4.13 (m, 3H), 3.81 (d, J=0.8 Hz,3H), 2.69 (dq, J=19.1, 2.8 Hz, 1H), 2.27-2.16 (m, 3H), 2.00-1.81 (m,2H), 1.26 (s, 12H), 1.22 (t, J=7.2 Hz, 3H). ¹⁹F NMR (376 MHz,CHLOROFORM-d) δ −60.84 (s, 3F). LC/MS m/z 481.13 (M+Na)⁺, 2.41 min (LCMSMethod 1).

Step 5. Preparation of ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((1-methyl-5-(trifluoromethyl)-1H-pyrazol-3-yl)oxy)methyl)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 88% yield as a solid, following theprocedure described in general procedure A step 5, using ethyl1-(((1-methyl-5-(trifluoromethyl)-1H-pyrazol-3-yl)oxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-ene-1-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 5.98 (s, 1H), 5.33 (br.s., 1H), 5.17 (d, J=4.8 Hz, 1H), 4.71 (s, 1H), 4.60 (s, 1H), 4.29-4.09(m, 4H), 3.80 (s, 3H), 3.12-3.00 (m, 8H), 2.79-2.46 (m, 6H), 2.24-0.88(m, 27H), 1.69 (s, 3H), 1.22 (t, J=7.0 Hz, 3H), 1.05 (s, 3H), 0.96 (s,3H), 0.96-0.89 (m, 6H), 0.85 (s, 3H). ¹⁹F NMR (376 MHz, CHLOROFORM-d)8-60.83 (s, 3F). LC/MS m/z 901.50 (M+H) 3.89 min (LCMS Method 2).

Step 6.4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((1-methyl-5-(trifluoromethyl)-1H-pyrazol-3-yl)oxy)methyl)cyclohex-3-ene-1-carboxylicacid was prepared in 56% yield as a solid, following the proceduredescribed in general procedure A step 6, using ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((1-methyl-5-(trifluoromethyl)-1H-pyrazol-3-yl)oxy)methyl)cyclohex-3-ene-1-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 6.00 (s, 1H), 5.35 (br.s., 1H), 5.19 (d, J=5.8 Hz, 1H), 4.71 (s, 1H), 4.60 (s, 1H), 4.34-4.21(m, 2H), 3.81 (s, 3H), 3.14-2.99 (m, 8H), 2.76-2.54 (m, 6H), 2.23-1.04(m, 27H), 1.69 (s, 3H), 1.08 (s, 3H), 0.97 (s, 3H), 0.97-0.92 (m, 6H),0.86 (s, 3H). ¹⁹F NMR (376 MHz, CHLOROFORM-d) δ −60.81 (s, 3F). LC/MSm/z 873.45 (M+H)⁺, 3.73 min (LCMS Method 2).

Example 4 Preparation of2-((1-carboxy-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-en-1-yl)methoxy)thiazole-4-carboxylicacid

Step 1. Preparation of ethyl2-((8-(ethoxycarbonyl)-1,4-dioxaspiro[4.5]decan-8-yl)methoxy)thiazole-4-carboxylate

The title compound was prepared as an oil without further purification,following the procedure described in general procedure A step 1-A, usingethyl 2-hydroxythiazole-4-carboxylate as reactant. LC/MS m/z 400.30(M+H)⁺, 2.18 min (LCMS Method 1).

Step 2. Preparation of ethyl2-((1-(ethoxycarbonyl)-4-oxocyclohexyl)methoxy)thiazole-4-carboxylate

The title compound was prepared in 26% yield (yield calculated over 2steps) as a solid, following the procedure described in generalprocedure A step 2, using crude ethyl 2-((8-(ethoxycarbonyl)-1,4-dioxaspiro[4.5]decan-8-yl)methoxy)thiazole-4-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.61 (s, 1H), 4.66 (s,2H), 4.38 (q, J=7.3 Hz, 2H), 4.26 (q, J=7.2 Hz, 2H), 2.58-2.48 (m, 4H),2.45-2.36 (m, 2H), 1.92-1.81 (m, 2H), 1.39 (t, J=7.2 Hz, 3H), 1.28 (t,J=7.2 Hz, 3H).

Step 3. Preparation of ethyl 2-((1-(ethoxycarbonyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-en-1-yl)methoxy)thiazole-4-carboxylate

The title compound was prepared in 40% yield as an oil, following theprocedure described in general procedure A step 3, using ethyl2-((1-(ethoxy carbonyl)-4-oxocyclohexyl)methoxy)thiazole-4-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.60 (s, 1H), 5.80-5.76(m, 1H), 4.66 (d, J=10.0 Hz, 1H), 4.60 (d, J=10.3 Hz, 1H), 4.38 (q,J=7.0 Hz, 2H), 4.19 (qd, J=7.1, 0.8 Hz, 2H), 2.87-2.79 (m, 1H),2.56-2.23 (m, 4H), 1.99-1.90 (m, 1H), 1.38 (t, J=7.2 Hz, 3H), 1.24 (t,J=7.0 Hz, 3H). ¹⁹F NMR (376 MHz, CHLOROFORM-d) δ −73.84 (s, 3F). LC/MSm/z 488.15 (M+H)⁺, 2.41 min (LCMS Method 1).

Step 4. Preparation of ethyl2-((1-(ethoxycarbonyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-en-1-yl)methoxy)thiazole-4-carboxylate

The title compound was prepared in 57% yield as an oil, following theprocedure described in general procedure A step 4, using ethyl2-((1-(ethoxycarbonyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-en-1-yl)methoxy)thiazole-4-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.58 (s, 1H), 6.54-6.49(m, 1H), 4.64 (d, J=10.0 Hz, 1H), 4.56 (d, J=10.0 Hz, 1H), 4.37 (q,J=7.0 Hz, 2H), 4.16 (q, J=7.2 Hz, 2H), 2.68 (dq, J=19.1, 3.0 Hz, 1H),2.27-2.16 (m, 3H), 2.00-1.81 (m, 2H), 1.38 (t, J=7.0 Hz, 3H), 1.26 (s,12H), 1.21 (t, J=7.0 Hz, 3H). LC/MS m/z 466.30 (M+H)⁺, 2.42 min (LCMSMethod 1).

Step 5. Preparation of ethyl2-((4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(ethoxycarbonyl)cyclohex-3-en-1-yl)methoxy)thiazole-4-carboxylate

The title compound was prepared in 79% yield as a solid, following theprocedure described in general procedure A step 5, using ethyl2-((1-(ethoxycarbonyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-en-1-yl)methoxy)thiazole-4-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.58 (s, 1H), 5.33 (br.s., 1H), 5.18 (d, J=6.0 Hz, 1H), 4.71 (d, J=2.0 Hz, 1H), 4.67-4.60 (m,2H), 4.59 (s, 1H), 4.37 (q, J=7.0 Hz, 2H), 4.20-4.09 (m, 2H), 3.12-2.96(m, 8H), 2.74-2.41 (m, 6H), 2.21-0.86 (m, 27H), 1.69 (s, 3H), 1.38 (t,J=1.2 Hz, 3H), 1.22 (t, J=7.2 Hz, 3H), 1.05 (s, 3H), 0.96 (s, 3H),0.96-0.90 (m, 6H), 0.85 (s, 3H). LC/MS m/z 908.60 (M+H)⁺, 3.05 min (LCMSMethod 3).

Step 6.2-((1-carboxy-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-en-1-yl)methoxy)thiazole-4-carboxylicacid was prepared in 85% yield as a solid, following the proceduredescribed in general procedure A step 6, using ethyl2-((4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(ethoxycarbonyl)cyclohex-3-en-1-yl)methoxy)thiazole-4-carboxylate as reactant.¹H NMR (500 MHz, METHANOL-d₄) δ 7.49 (s, 1H), 5.34 (br. s., 1H), 5.21(d, J=4.7 Hz, 1H), 4.78 (s, 1H), 4.68 (s, 1H), 4.61-4.53 (m, 2H),3.27-3.06 (m, 11H), 2.99-2.96 (m, 1H), 2.89-2.80 (m, 1H), 2.67-2.58 (m,1H), 2.35-1.04 (m, 27H), 1.73 (s, 3H), 1.18 (s, 3H), 1.09 (s, 3H),1.00-0.96 (m, 6H), 0.92 (s, 3H). LC/MS m/z 852.50 (M+H)⁺, 2.86 min (LCMSMethod 3).

Example 5

Preparation of4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((4-methyl-1,2,5-thiadiazol-3-yl)oxy)methyl)cyclohex-3-ene-1-carboxylicacid.

Step 1. Preparation of ethyl8-(((4-methyl-1,2,5-thiadiazol-3-yl)oxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate

The title compound was prepared in 64% yield as an oil, following theprocedure described in general procedure A step 1-A, using4-methyl-1,2,5-thiadiazol-3-ol as reactant. ¹H NMR (400 MHz,CHLOROFORM-d) δ 4.43 (s, 2H), 4.18 (q, J=7.1 Hz, 2H), 3.99-3.92 (m, 4H),2.36 (s, 3H), 2.31-2.24 (m, 2H), 1.75-1.66 (m, 6H), 1.24 (t, J=7.2 Hz,3H). LC/MS m/z 343.20 (M+H)⁺, 2.17 min (LCMS Method 1).

Step 2. Preparation of ethyl1-(((4-methyl-1,2,5-thiadiazol-3-yl)oxy)methyl)-4-oxocyclohexane-1-carboxylate

The title compound was prepared in 81% yield as an oil, following theprocedure described in general procedure A step 2, using ethyl8-(((4-methyl-1,2,5-thiadiazol-3-yl)oxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 4.51 (s, 2H), 4.26 (q,J=7.1 Hz, 2H), 2.61-2.38 (m, 6H), 2.37 (s, 3H), 1.91-1.82 (m, 2H), 1.28(t, J=7.2 Hz, 3H). LC/MS m/z 299.20 (M+H)⁺, 1.94 min (LCMS Method 1).

Step 3. Preparation of ethyl1-(((4-methyl-1,2,5-thiadiazol-3-yl)oxy)methyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 60% yield as an oil, following theprocedure described in general procedure A step 3, using ethyl1-(((4-methyl-1,2,5-thiadiazol-3-yl)oxy)methyl)-4-oxocyclohexane-1-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 5.82-5.78 (m, 1H), 4.52(d, J=10.3 Hz, 1H), 4.47 (d, J=10.3 Hz, 1H), 4.26-4.12 (m, 2H),2.90-2.82 (m, 1H), 2.59-2.27 (m, 4H), 2.36 (s, 3H), 2.00-1.93 (m, 1H),1.24 (t, J=7.0 Hz, 3H). ¹⁹F NMR (376 MHz, CHLOROFORM-d) 8-73.83 (s, 3F).LC/MS m/z 431.15 (M+H)⁺, 2.41 min (LCMS Method 1).

Step 4. Preparation of ethyl1-(((4-methyl-1,2,5-thiadiazol-3-yl)oxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 74% yield as an oil, following theprocedure described in general procedure A step 4, using ethyl1-(((4-methyl-1,2,5-thiadiazol-3-yl)oxy)methyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-ene-1-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 6.56-6.52 (m, 1H), 4.51(d, J=10.0 Hz, 1H), 4.44 (d, J=10.0 Hz, 1H), 4.16 (qd, J=7.1, 1.1 Hz,2H), 2.71 (dq, J=19.1, 3.3 Hz, 1H), 2.35 (s, 3H), 2.31-2.17 (m, 3H),2.04-1.85 (m, 2H), 1.26 (s 12H), 1.21 (t, J=7.0 Hz, 3H). LC/MS m/z409.25 (M+H)⁺, 2.45 min (LCMS Method 1).

Step 5. Preparation of ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((4-methyl-1,2,5-thiadiazol-3-yl)oxy)methyl)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 73% yield as a solid, following theprocedure described in general procedure A step 5, using ethyl1-(((4-methyl-1,2,5-thiadiazol-3-yl)oxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-ene-1-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 5.36 (br. s., 1H), 5.19(d, J=5.3 Hz, 1H), 4.76 (s, 1H), 4.63 (s, 1H), 4.56-4.44 (m, 2H),4.21-4.09 (m, 2H), 3.17-3.00 (m, 8H), 2.98-2.59 (m, 6H), 2.23-0.82 (m,27H), 2.35 (s, 3H), 1.70 (s, 3H), 1.22 (t, J=1.2 Hz, 3H), 1.06 (s, 3H),0.98 (s, 3H), 0.97-0.91 (m, 6H), 0.85 (s, 3H). LC/MS m/z 851.55 (M+H)⁺,3.07 mm (LCMS Method 3).

Step 6.4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((4-methyl-1,2,5-thiadiazol-3-yl)oxy)methyl)cyclohex-3-ene-1-carboxylicacid was prepared in 53% yield as a solid, following the proceduredescribed in general procedure A step 6, using ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((4-methyl-1,2,5-thiadiazol-3-yl)oxy)methyl)cyclohex-3-ene-1-carboxylateas reactant.

¹H NMR (400 MHz, CHLOROFORM-d) δ 5.36 (br. s., 1H), 5.18 (br. s., 1H),4.74 (br. s., 1H), 4.65 (br. s., 1H), 4.59-4.45 (m, 2H), 3.24-2.98 (m,9H), 2.89-2.51 (m, 5H), 2.34 (s, 3H), 1.68 (s, 3H), 2.22-0.97 (m, 27H),1.15 (s, 3H), 1.02 (s, 3H), 0.97-0.89 (m, 6H), 0.86 (s, 3H). LC/MS m/z823.55 (M+H)⁺, 2.85 min (LCMS Method 3).

Example 6 Preparation of1-(((1,2,5-thiadiazol-3-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylicacid

Step 1. Preparation of ethyl8-(((1,2,5-thiadiazol-3-yl)oxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate

The title compound was prepared in 92% yield as an oil, following theprocedure described in general procedure A step 1-A, using1,2,5-thiadiazol-3-ol as reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.97(s, 1H), 4.46 (s, 2H), 4.18 (q, J=7.1 Hz, 2H), 4.00-3.92 (m, 4H),2.32-2.21 (m, 2H), 1.76-1.66 (m, 6H), 1.24 (t, J=1.2 Hz, 3H). LC/MS m/z329.20 (M+H)⁺, 2.07 mm (LCMS Method 1).

Step 2. Preparation of ethyl1-(((1,2,5-thiadiazol-3-yl)oxy)methyl)-4-oxocyclohexane-1-carboxylate

The title compound was prepared in 80% yield as an oil, following theprocedure described in general procedure A step 2, using ethyl8-(((1,2,5-thiadiazol-3-yl)oxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.99 (s, 1H), 4.54 (s,2H), 4.26 (q, J=7.0 Hz, 2H), 2.60-2.50 (m, 4H), 2.47-2.38 (m, 2H),1.93-1.82 (m, 2H), 1.28 (t, J=7.0 Hz, 3H). LC/MS m/z 285.15 (M+H)⁺, 1.85min (LCMS Method 1).

Step 3. Preparation of ethyl1-(((1,2,5-thiadiazol-3-yl)oxy)methyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 34% yield as an oil, following theprocedure described in general procedure A step 3, using ethyl1-(((1,2,5-thiadiazol-3-yl)oxy)methyl)-4-oxocyclohexane-1-carboxylate asreactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.99 (s, 1H), 5.81-5.78 (m,1H), 4.55 (d, J=10.3 Hz, 1H), 4.50 (d, J=10.3 Hz, 1H), 4.20 (qd, J=7.1,0.8 Hz, 2H), 2.90-2.81 (m, 1H), 2.57-2.25 (m, 4H), 2.04-1.95 (m, 1H),1.24 (t, J=7.2 Hz, 3H). ¹⁹F NMR (376 MHz, CHLOROFORM-d) δ −73.83 (s,3F). LC/MS m/z 417.10 (M+H)⁺, 2.37 mm (LCMS Method 1).

Step 4. Preparation of ethyl1-(((1,2,5-thiadiazol-3-yl)oxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 69% yield as an oil, following theprocedure described in general procedure A step 4, using ethyl1-(((1,2,5-thiadiazol-3-yl)oxy)methyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-ene-1-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.96 (s, 1H), 6.56-6.52(m, 1H), 4.54 (d, J=10.0 Hz, 1H), 4.45 (d, J=10.0 Hz, 1H), 4.16 (q,J=7.0 Hz, 2H), 2.71 (dq, J=18.9, 3.4 Hz, 1H), 2.30-2.17 (m, 3H),2.03-1.94 (m, 1H), 1.92-1.83 (m, 1H), 1.26 (s, 12H), 1.21 (t, J=7.2 Hz,3H). LC/MS m/z 395.30 (M+H)⁺, 2.40 min (LCMS Method 1).

Step 5. Preparation of ethyl1-(((1,2,5-thiadiazol-3-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 76% yield as a solid, following theprocedure described in general procedure A step 5, using ethyl1-(((1,2,5-thiadiazol-3-yl)oxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-ene-1-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.97 (s, 1H), 5.35 (br.s., 1H), 5.18 (d, J=5.0 Hz, 1H), 4.71 (s, 1H), 4.59 (s, 1H), 4.58-4.49(m, 2H), 4.16 (q, J=7.5 Hz, 2H), 3.13-2.98 (m, 8H), 2.76-2.43 (m, 6H),2.22-0.82 (m, 27H), 1.69 (s, 3H), 1.22 (t, J=7.2 Hz 3H), 1.06 (s, 3H),0.97 (s, 3H), 0.96-0.91 (m, 6H), 0.85 (s, 3H). LC/MS m/z 837.55 (M+H)⁺,3.08 min (LCMS Method 3).

Step 6.1-(((1,2,5-thiadiazol-3-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylicacid was prepared in 56% yield as a solid, following the proceduredescribed in general procedure A step 6, using ethyl1-(((1,2,5-thiadiazol-3-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.98 (s, 1H), 5.37 (br.s., 1H), 5.19 (br. s., 1H), 4.76 (s, 1H), 4.66 (s, 1H), 4.62-4.49 (m,2H), 3.23-3.00 (m, 8H), 2.90-2.53 (m, 6H), 2.28-0.89 (m, 27H), 1.69 (s,3H), 1.16 (s, 3H), 1.03 (s, 3H), 0.97-0.91 (m, 6H), 0.86 (s, 3H). LC/MSm/z 809.50 (M+H)⁺, 2.90 min (LCMS Method 3).

Example 7 Preparation of4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((1-ethyl-1H-1,2,3-triazol-5-yl)oxy)methyl)cyclohex-3-ene-1-carboxylicacid

Step 1. Preparation of ethyl8-(((1-ethyl-1H-1,2,3-triazol-5-yl)oxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate

The title compound was prepared in 56% yield as an oil, following theprocedure described in general procedure A step 1-B, using1-ethyl-1H-1,2,3-triazol-5-ol as reactant. 10 ¹H NMR (500 MHz,CHLOROFORM-d) δ 7.07 (s, 1H), 4.19 (q, J=7.1 Hz, 3H), 4.17 (q, J=7.3 Hz,2H), 4.10 (s, 2H), 4.01-3.92 (m, 4H), 2.33-2.24 (m, 2H), 1.76-1.61 (m,6H), 1.44 (t, J=7.3 Hz, 3H), 1.25 (t, J=7.2 Hz, 3H). LC/MS m/z 340.25(M+Na) 1.91 min (LCMS Method 1).

Step 2. Preparation of ethyl1-(((1-ethyl-1H-1,2,3-triazol-5-yl)oxy)methyl)-4-oxocyclohexane-1-carboxylate

The title compound was prepared in 86% yield as an oil, following theprocedure described in general procedure A step 2, using ethyl8-(((1-ethyl-1H-1,2,3-triazol-5-yl)oxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.09 (s, 1H), 4.26 (q,J=7.0 Hz, 2H), 4.18 (q, J=7.5 Hz, 2H), 4.17 (s, 2H), 2.61-2.50 (m, 4H),2.47-2.38 (m, 2H), 1.89-1.78 (m, 2H), 1.45 (t, J=7.4 Hz, 3H), 1.28 (t,J=7.2 Hz, 3H). LC/MS m/z 296.25 (M+H)⁺, 1.62 min (LCMS Method 1).

Step 3. Preparation of ethyl1-(((1-ethyl-1H-1,2,3-triazol-5-yl)oxy)methyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 35% yield as an oil, following theprocedure described in general procedure A step 3, using ethyl1-(((1-ethyl-1H-1,2,3-triazol-5-yl)oxy)methyl)-4-oxocyclohexane-1-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.09 (s, 1H), 5.82-5.78(m, 1H), 4.24-4.13 (m, 6H), 2.90-2.81 (m, 1H), 2.60-2.24 (m, 4H),2.00-1.92 (m, 1H), 1.44 (t, J=7.3 Hz, 3H), 1.25 (t, J=7.2 Hz, 3H). ¹⁹FNMR (376 MHz, CHLOROFORM-d) 8-73.82 (s, 3F). LC/MS m/z 428.20 (M+H)⁺,2.15 min (LCMS Method 1).

Step 4. Preparation of ethyl1-(((1-ethyl-1H-1,2,3-triazol-5-yl)oxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 57% yield as an oil, following theprocedure described in general procedure A step 4, using ethyl1-(((1-ethyl-1H-1,2,3-triazol-5-yl)oxy)methyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-ene-1-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.07 (s, 1H), 6.55-6.50(m, 1H), 4.21-4.11 (m, 6H), 2.69 (dq, J=19.0, 2.9 Hz, 1H), 2.31-2.09 (m,3H), 2.02-1.85 (m, 2H), 1.43 (t, J=7.3 Hz, 3H), 1.27 (s, 12H), 1.22 (t,J=7.2 Hz, 3H). LC/MS m/z 406.20 (M+H)⁺, 2.22 min (LCMS Method 1).

Step 5. Preparation of ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((1-ethyl-1H-1,2,3-triazol-5-yl)oxy)methyl)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 90% yield as a solid, following theprocedure described in general procedure A step 5, using ethyl1-(((1-ethyl-1H-1,2,3-triazol-5-yl)oxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-ene-1-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.07 (s, 1H), 5.35 (br.s., 1H), 5.18 (d, J=6.0 Hz, 1H), 4.70 (s, 1H), 4.59 (s, 1H), 4.23-4.11(m, 6H), 3.11-2.97 (m, 8H), 2.71-2.42 (m, 6H), 2.24-0.86 (m, 27H), 1.68(s, 3H), 1.42 (t, J=7.3 Hz, 3H), 1.23 (t, J=7.0 Hz, 3H), 1.05 (s, 3H),0.96 (s, 3H), 0.95-0.90 (m, 6H), 0.85 (s, 3H). LC/MS m/z 848.60 (M+H)⁺,2.74 min (LCMS Method 3).

Step 6.4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorphoIino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((1-ethyl-1H-1,2,3-triazol-5-yl)oxy)methyl)cyclohex-3-ene-1-carboxylicacid was prepared in 61% yield as a solid, following the proceduredescribed in general procedure A step 6, using ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((1-ethyl-1H-1,2,3-triazol-5-yl)oxy)methyl)cyclohex-3-ene-1-carboxylateas reactant.

¹H NMR (400 MHz, CHLOROFORM-d) δ 7.10 (s, 1H), 5.36 (br. s., 1H), 5.18(br. s., 1H), 4.69 (s, 1H), 4.59 (s, 1H), 4.28-4.20 (m, 2H), 4.16 (q,J=7.3 Hz, 2H), 3.13-2.99 (m, 8H), 2.82-2.55 (m, 6H), 2.24-1.00 (m, 27H),1.68 (s, 3H), 1.43 (t, J=7.3 Hz, 3H), 1.09 (s, 3H), 0.98 (s, 3H),0.96-0.91 (m, 6H), 0.85 (s, 3H). LC/MS m/z 820.55 (M+H)⁺, 2.86 min (LCMSMethod 3).

Example 8 Preparation of1-((benzo[d]isothiazol-3-yloxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylicacid

Step 1. Preparation of ethyl8-((benzo[d]isothiazol-3-yloxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate

The title compound was prepared in 26% yield as an oil, following theprocedure described in general procedure A step 1-B, usingbenzo[d]isothiazol-3(2H)-one as reactant. 10 ¹H NMR (500 MHz,CHLOROFORM-d) δ 7.88 (dd, J=8.1, 0.9 Hz, 1H), 7.78 (d, J=8.1 Hz, 1H),7.53 (ddd, J=8.2, 7.1, 1.1 Hz, 1H), 7.39 (td, J=7.5, 0.8 Hz, 1H),4.63-4.59 (m, 2H), 4.20 (q, J=7.1 Hz, 2H), 3.97 (t, J=2.6 Hz, 4H),2.41-2.31 (m, 2H), 1.82-1.73 (m, 6H), 1.23 (t, J=1.1 Hz, 3H). LC/MS m/z378.25 (M+H)⁺, 4.17 min (LCMS Method 4).

Step 2. Preparation of ethyl1-((benzo[d]isothiazol-3-yloxy)methyl)-4-oxocyclohexane-1-carboxylate

The title compound was prepared in 88% yield as a wax, following theprocedure described in general procedure A step 2, using ethyl8-((benzo[d]isothiazol-3-yloxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.87 (dt, J=8.0, 1.0 Hz,1H), 7.79 (dt, J=8.2, 0.8 Hz, 1H), 7.54 (ddd, J=8.2, 7.0, 1.1 Hz, 1H),7.40 (ddd, J=8.0, 7.0, 1.0 Hz, 1H), 4.68 (s, 2H), 4.26 (q, J=7.3 Hz,2H), 2.66-2.51 (m, 4H), 2.49-2.40 (m, 2H), 1.99-1.88 (m, 2H), 1.26 (t,J=7.2 Hz, 3H). LC/MS m/z 334.20 (M+H)⁺, 2.31 min (LCMS Method 1).

Step 3. Preparation of ethyl1-((benzo[d]isothiazol-3-yloxy)methyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 64% yield as an oil, following theprocedure described in general procedure A step 3, using ethyl1-((benzo[d]isothiazol-3-yloxy)methyl)-4-oxocyclohexane-1-carboxylate asreactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.86 (dt, J=8.0, 1.0 Hz, 1H),7.79 (dt, J=8.1, 0.8 Hz, 1H), 7.54 (ddd, J=8.2, 7.0, 1.1 Hz, 1H),7.43-7.39 (m, 1H), 5.83-5.79 (m, 1H), 4.68 (d, J=10.0 Hz, 1H), 4.63 (d,J=10.3 Hz, 1H), 4.20 (qd, J=7.2, 2.1 Hz, 2H), 2.97-2.88 (m, 1H),2.59-2.32 (m, 4H), 2.07-1.98 (m, 1H), 1.23 (t, J=1.2 Hz, 3H). ¹⁹F NMR(376 MHz, CHLOROFORM-d) 8-73.83 (s, 3F). LC/MS m/z 466.15 (M+H)⁺, 2.51min (LCMS Method 1).

Step 4. Preparation of ethyl1-((benzo[d]isothiazol-3-yloxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 61% yield as an oil, following theprocedure described in general procedure A step 4, using ethyl1-((benzo[d]isothiazol-3-yloxy)methyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-ene-1-carboxylateas reactant.

¹H NMR (400 MHz, CHLOROFORM-d) δ 7.85 (dt, J=7.9, 0.9 Hz, 1H), 7.76 (dt,J=8.2, 0.8 Hz, 1H), 7.52 (ddd, J=8.2, 7.0, 1.1 Hz, 1H), 7.42-7.37 (m,1H), 6.58-6.54 (m. 1H), 4.66 (d, J=10.0 Hz, 1H), 4.58 (d, J=10.0 Hz,1H), 4.16 (qd, J=7.1, 1.0 Hz, 2H), 2.76 (dq, J=18.9, 2.7 Hz, 1H),2.37-2.28 (m, 1H), 2.27-2.20 (m, 2H), 2.07-1.89 (m, 2H), 1.28-1.25 (m,12H), 1.19 (t, J=7.2 Hz, 3H). LC/MS m/z 444.25 (M+H)⁺, 2.58 min (LCMSMethod 1).

Step 5. Preparation of ethyl1-((benzo[d]isothiazol-3-yloxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 47% yield as a solid, following theprocedure described in general procedure A step 5, using ethyl1-((benzo[d]isothiazol-3-yloxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-ene-1-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.87 (d, J=7.8 Hz, 1H),7.77 (d, J=8.0 Hz, 1H), 7.52 (td, J=7.6, 1.1 Hz, 1H), 7.37 (td, J=7.5,1.0 Hz, 1H), 5.37 (br. s., 1H), 5.20 (d, J=6.0 Hz, 1H), 4.76 (s, 1H),4.64 (s, 1H), 4.19-4.16 (m, 2H), 4.13 (q, J=7.1 Hz, 2H), 3.17-3.03 (m,8H), 2.79-2.36 (m, 6H), 2.28-0.83 (27H), 1.70 (s, 3H), 1.27 (t, J=7.3Hz, 3H), 1.08 (s, 3H), 0.99 (s, 3H), 0.99-0.95 (m, 6H), 0.85 (s, 3H).LC/MS m/z 886.55 (M+H)⁺, 3.07 min (LCMS Method 3).

Step 6.1-((benzo[d]isothiazol-3-yloxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylicacid was prepared in 21% yield as a solid, following the proceduredescribed in general procedure A step 6, using ethyl1-((benzo[d]isothiazol-3-yloxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.88 (d, J=8.0 Hz, 1H),7.77 (d, J=8.3 Hz, 1H), 7.55-7.49 (m, 1H), 7.40-7.33 (m, 1H), 5.39 (br.s., 1H), 5.20 (br. s., 1H), 4.78 (s, 1H), 4.71 (s, 1H), 4.74-4.64 (m,2H), 3.34-2.52 (m, 14H), 2.33-1.00 (m, 27H), 1.69 (s, 3H), 1.15 (s, 3H),1.04 (s, 3H), 0.98-0.91 (m, 6H), 0.87 (s, 3H). LC/MS m/z 858.50 (M+H)⁺,2.88 min (LCMS Method 3).

Example 9 Preparation of4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-4-yloxy)methyl)cyclohex-3-ene-1-carboxylicacid

Step 1. Preparation of ethyl8-((pyridin-4-yloxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate

The title compound was prepared in 77% yield, following the proceduredescribed in general procedure A step 1-A, using 4-hydroxypyridine asreactant. ¹H NMR (500 MHz, CHLOROFORM-d) δ 8.46-8.41 (m, 2H), 6.81-6.77(m, 2H), 4.20 (q, J=1.2 Hz, 2H), 4.04 (s, 2H), 4.01-3.93 (m, 4H),2.37-2.25 (m, 2H), 1.80-1.66 (m, 6H), 1.24 (t, J=7.1 Hz, 3H). LC/MS: m/e322.05 (M+H)⁺, 2.26 min (LCMS Method 11).

Step 2. Preparation of ethyl4-oxo-1-((pyridin-4-yloxy)methyl)cyclohexane-1-carboxylate

The title compound was prepared in 64% yield, following the proceduredescribed in general procedure A step 2, using ethyl8-((pyridin-4-yloxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate asreactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 8.27-8.23 (m, 2H), 6.67-6.62(m, 2H), 3.97 (s, 2H), 3.92 (q, J=7.3 Hz, 2H), 2.43-2.31 (m, 4H),2.27-2.17 (m, 2H), 1.77-1.66 (m, 2H), 1.07 (t, J=7.3 Hz, 3H). LC/MS: m/e278.05 (M+H)⁺, 0.81 min (LCMS Method 8).

Step 3. Preparation of ethyl1-((pyridin-4-yloxy)methyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 66% yield, following the proceduredescribed in general procedure A step 3, using ethyl4-oxo-1-((pyridin-4-yloxy)methyl)cyclohexanecarboxylate as reactant. ¹HNMR (400 MHz, CHLOROFORM-d) δ 8.47-8.44 (m, 2H), 6.83-6.79 (m, 2H), 5.82(t, J=4.1 Hz, 1H), 4.21 (q, J=7.2 Hz, 2H), 4.17-4.07 (m, 2H), 2.91-2.82(m, 1H), 2.59-2.47 (m, 1H), 2.45-2.25 (m, 4H), 2.09-2.00 (m, 2H), 1.25(t, J=7.2 Hz, 3H). LC/MS: m/e 410.00 (M+H)⁺, 1.92 min (LCMS Method 8).

Step 4. Preparation of ethyl1-((pyridin-4-yloxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 59% yield, following the proceduredescribed in general procedure A step 4, using ethyl1-((pyridin-4-yloxy)methyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-enecarboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 8.33-8.30 (m, 2H),6.73-6.69 (m, 2H), 6.46 (br. s., 1H), 4.10-4.04 (m, 2H), 4.02-3.95 (m,2H), 2.63 (dd, J=19.2, 2.9 Hz, 1H), 2.23-2.12 (m, 2H), 2.12-2.01 (m,1H), 1.94-1.77 (m, 2H), 1.17 (s, 12H), 1.11 (t, J=7.2 Hz, 3H). LC/MS:m/e 388.10 (M+H)⁺, 1.90 min (LCMS Method 8).

Step 5. Preparation of ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-4-yloxy)methyl)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 34% yield, following the proceduredescribed in general procedure A step 5, using ethyl1-((pyridin-4-yloxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-enecarboxylateas reactant. LC/MS: m/e 831.45 (M+H)⁺, 2.54 min (LCMS Method 3). ¹H NMR(400 MHz, CHLOROFORM-d) δ 8.42 (d, J=6.0 Hz, 2H), 6.80 (d, J=6.3 Hz,2H), 5.36 (br. s., 1H), 5.18 (d, J=4.8 Hz, 1H), 4.71 (d, J=1.8 Hz, 1H),4.59 (s, 1H), 4.20-4.06 (m, 4H), 3.12-2.97 (m, 8H), 2.74-2.51 (m, 4H),2.51-2.40 (m, 1H), 2.31-2.12 (m, 4H), 2.11-1.98 (m, 3H), 1.98-1.80 (m,5H), 1.80-1.62 (m, 2H), 1.69 (s, 3H), 1.62-1.37 (m, 10H), 1.37-1.17 (m,4H), 1.26 (t, J=7.2 Hz, 3H), 1.16-0.99 (m, 3H), 0.99-0.93 (m, 6H),0.93-0.87 (m, 3H), 0.87-0.81 (m, 3H).

Step 6.4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-4-yloxy)methyl)cyclohex-3-ene-1-carboxylicacid was prepared in 47% yield, following the procedure described ingeneral procedure A step 6, using ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-4-yloxy)methyl)cyclohex-3-enecarboxylateas reactant. ¹H NMR (400 MHz, METHANOL-d₄) δ 8.33 (d, J=6.3 Hz, 2H),6.98 (d, J=5.5 Hz, 2H), 5.38 (br. s., 1H), 5.21 (d, J=5.0 Hz, 1H),4.80-4.71 (m, 1H), 4.65 (s, 1H), 4.28-4.12 (m, 2H), 3.24-3.00 (m, 8H),2.94-2.72 (m, 5H), 2.66 (d, J=18.1 Hz, 1H), 2.37-1.97 (m, 8H), 1.97-1.78(m, 1H), 1.72 (s, 3H), 1.78-1.69 (m, 3H), 1.66-1.21 (m, 14H), 1.16 (s,3H), 1.20-1.08 (m, 2H), 1.05 (s, 3H), 0.99 (s, 3H), 0.96 (s, 3H), 0.91(s, 3H). LC/MS: m/e 802.45 (M+H)⁺, 2.50 min (LCMS Method 3).

Example 10 Preparation of4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-3-yloxy)methyl)cyclohex-3-enecarboxylicacid

Step 1. Preparation of ethyl8-((pyridin-3-yloxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate

The title compound was prepared in 84% yield, following the proceduredescribed in general procedure A step 1-A, using 3-hydroxypyridine asreactant ¹H NMR (500 MHz, CHLOROFORM-d) δ 8.30 (d, J=2.7 Hz, 1H), 8.24(dd, J=4.4, 1.4 Hz, 1H), 7.25-7.15 (m, 2H), 4.21 (q, J=7.2 Hz, 2H), 4.05(s, 2H), 4.00-3.93 (m, 4H), 2.38-2.25 (m, 2H), 1.83-1.66 (m, 6H),1.32-1.22 (m, 3H). LC/MS: m/e 322.10 (M+H)⁺, 2.534 min (LCMS Method 11).

Step 2. Preparation of ethyl4-oxo-1-((pyridin-3-yloxy)methyl)cyclohexane-1-carboxylate

The title compound was prepared in 47.8% yield, following the proceduredescribed in general procedure A step 2, using ethyl8-((pyridin-3-yloxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate asreactant. LCMS: m/e 279.00 (M+H)⁺, 2.079 min (LCMS Method 8). ¹H NMR(400 MHz, CHLOROFORM-d) δ 8.22 (dd, J=2.6, 0.9 Hz, 1H), 8.16 (dd, J=4.3,1.8 Hz, 1H), 7.19-7.09 (m, 2H), 4.18 (q, J=7.0 Hz, 2H), 4.07-4.04 (m,2H), 2.55-2.41 (m, 4H), 2.38-2.28 (m, 2H), 1.88-1.75 (m, 2H), 1.19 (t,J=7.2 Hz, 3H).

Step 3. Preparation of ethyl1-((pyridin-3-yloxy)methyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 51.9% yield, following the proceduredescribed in general procedure A step 3, using ethyl4-oxo-1-((pyridin-3-yloxy)methyl)cyclohexanecarboxylate as reactant.LCMS: m/e 410.00 (M+H)⁺, 1.983 min (LCMS Method 8). ¹H NMR (400 MHz,CHLOROFORM-d) δ 8.26-8.21 (m, 1H), 8.19-8.13 (m, 1H), 7.20-7.09 (m, 2H),5.78-5.70 (m, 1H), 4.16-4.10 (m, 2H), 4.09-4.02 (m, 2H), 2.84-2.74 (m,1H), 2.52-2.40 (m, 1H), 2.38-2.26 (m, 2H), 2.26-2.17 (m, 1H), 2.01-1.92(m, 1H), 1.21-1.15 (m, 3H).

Step 4. Preparation of ethyl1-((pyridin-3-yloxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 88% yield, following the proceduredescribed in general procedure A step 4, using ethyl1-((pyridin-3-yloxy)methyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-enecarboxylateas reactant. LCMS: m/e 388.10 (M+H)⁺, 1.986 min (LCMS Method 8). ¹H NMR(400 MHz, CHLOROFORM-d) δ 8.20 (dd, J=2.4, 0.9 Hz, 1H), 8.13 (dd, J=4.0,2.0 Hz, 1H), 7.18-7.10 (m, 2H), 6.52-6.45 (m, 1H), 4.16-3.95 (m, 4H),2.71-2.60 (m, 1H), 2.26-2.03 (m, 3H), 1.96-1.79 (m, 2H), 1.20-1.18 (m,12H), 1.14 (t, J=7.2 Hz, 3H).

Step 5. Preparation of ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-3-yloxy)methyl)cyclohex-3-ene-1-carboxylateand ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-5a,5b,8,8,11a-pentamethyl-3a-((2-(4-(methylsulfonyl)piperidin-1-yl)ethyl)amino)-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-3-yloxy)methyl)cyclohex-3-enecarboxylate

The title compounds were prepared in 26.4% and 28.4 yields respectively,following the procedure described in general procedure A step 5, usingethyl1-((pyridin-3-yloxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-enecarboxylateas reactant.

For ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-3-yloxy)methyl)cyclohex-3-ene-1-carboxylate:LCMS: m/e 830.50 (M+H)⁺, 2.363 min (LCMS Method 8). ¹H NMR (400 MHz,CHLOROFORM-d) δ 8.28 (d, J=2.0 Hz, 1H), 8.21 (dd, J=4.0, 2.0 Hz, 1H),7.23-7.16 (m, 2H), 5.35 (br. s., 1H), 5.17 (d, J=4.8 Hz, 1H), 4.72 (d,J=1.8 Hz, 1H), 4.59 (s, 1H), 4.15-4.09 (m, 4H), 3.14-2.96 (m, 8H),2.91-2.48 (m, 6H), 1.68 (s, 3H), 1.05 (s, 3H), 2.29-1.00 (m, 30H),0.97-0.89 (m, 9H), 0.86-0.81 (m, 3H).

For ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-5a,5b,8,8,11a-pentamethyl-3a-((2-(4-(methylsulfonyl)piperidin-1-yl)ethyl)amino)-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-3-yloxy)methyl)cyclohex-3-enecarboxylate:LCMS: m/e 858.55 (M+H)⁺, 2.454 min (LCMS Method 8). ¹H NMR (400 MHz,CHLOROFORM-d) δ 8.28 (d, J=1.8 Hz, 1H), 8.21 (dd, J=4.0, 1.8 Hz, 1H),7.20-7.16 (m, 2H), 5.35 (br. s., 1H), 5.17 (d, J=4.8 Hz, 1H), 4.72 (d,J=1.8 Hz, 1H), 4.58 (s, 1H), 4.20-4.05 (m, 4H), 3.11 (t, J=8.5 Hz, 2H),2.83 (s, 3H), 2.88-2.76 (m, 1H), 2.2.74-2.38 (m, 7H), 1.68 (s, 3H), 1.06(s, 3H), 2.27-0.78 (m, 47H).

Step 6.4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-3-yloxy)methyl)cyclohex-3-enecarboxylicacid was prepared in 68.1% yield, following the procedure described ingeneral procedure A step 6, using ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-3-yloxy)methyl)cyclohex-3-enecarboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 8.32 (s, 1H), 8.21 (br.s., 1H), 7.23 (br. s., 2H), 5.37 (br. s., 1H), 5.18 (br. s., 1H), 4.71(s, 1H), 4.60 (s, 1H), 4.23-4.08 (m, 2H), 3.16-2.99 (m, 8H), 2.89-2.57(m, 6H), 2.33-1.79 (m, 9H), 1.68 (s, 3H), 1.11 (s, 3H), 0.99 (s, 3H),0.96 (s, 3H), 0.93-0.92 (m, 3H), 0.86 (s, 3H), 1.75-0.81 (m, 18H).LC/MS: m/e 802.45 (M+H)⁺, 2.346 min (LCMS Method 8).

Example 11 Preparation of4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-5a,5b,8,8,11a-pentamethyl-3a-((2-(4-(methylsulfonyl)piperidin-1-yl)ethyl)amino)-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-3-yloxy)methyl)cyclohex-3-enecarboxylicacid

The title compound was prepared in 4.02% yield, following the proceduredescribed in general procedure A step 6, using ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-5a,5b,8,8,11a-pentamethyl-3a-((2-(4-(methylsulfonyl)piperidin-1-yl)ethyl)amino)-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-3-yloxy)methyl)cyclohex-3-enecarboxylateas reactant. LCMS: m/e 830.50 (M+H)⁺, 2.367 min (LCMS Method 8). ¹H NMR(400 MHz, CHLOROFORM-d) δ 8.31 (s, 1H), 8.20 (t, J=2.9 Hz, 1H), 7.22 (d,J=2.3 Hz, 2H), 5.37 (br. s., 1H), 5.18 (d, J=5.5 Hz, 1H), 4.76-4.67 (m,1H), 4.59 (s, 1H), 4.16 (br. s., 2H), 3.13 (t, J=10.2 Hz, 2H), 2.92-2.59(m, 9H), 2.48 (d, J=11.5 Hz, 1H), 2.31-1.78 (m, 15H), 1.68 (s, 3H), 1.12(s, 3H), 0.98 (s, 3H), 0.96 (s, 3H), 0.93-0.92 (m, 3H), 0.85 (s, 3H),1.71-0.77 (m, 18H).

Example 12 Preparation of4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((5-methylisothiazol-3-yl)oxy)methyl)cyclohex-3-ene-1-carboxylicacid

Step 1. Preparation of ethyl8-((isothiazol-3-yloxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate

The title compound was prepared in 69% yield, following the proceduredescribed in general procedure A step 1-A, using 5-methylisothiazol-3-olas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 6.32 (d, J=1.0 Hz, 1H),4.37 (br. s, 2H), 4.17 (q, J=7.0 Hz, 2H), 3.95 (s, 4H), 2.47 (d, J=1.0Hz, 3H), 2.28-2.21 (m, 2H), 1.76-1.63 (m, 6H), 1.23 (t, J=7.2 Hz, 3H).LC/MS: m/e 342.10 (M+H)⁺, 3.67 min (LCMS Method 11).

Step 2. Preparation of ethyl1-(((5-methylisothiazol-3-yl)oxy)methyl)-4-oxocyclohexane-1-carboxylate

The title compound was prepared in 87% yield, following the proceduredescribed in general procedure A step 2, using ethyl8-(((5-methylisothiazol-3-yl)oxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 6.34 (s, 1H), 4.48 (s,2H), 4.26 (q, J=7.1 Hz, 2H), 2.59-2.45 (m, 4H), 2.45-2.36 (m, 2H),1.94-1.82 (m, 2H), 1.29 (t, J=7.1 Hz, 3H). LC/MS: m/e 298.05 (M+H)⁺,2.20 min (LCMS Method 8).

Step 3. Preparation of ethyl1-(((5-methylisothiazol-3-yl)oxy)methyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 87% yield, following the proceduredescribed in general procedure A step 3, using ethyl1-(((5-methylisothiazol-3-yl)oxy)methyl)-4-oxocyclohexanecarboxylate asreactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 6.31 (s, 1H), 5.74 (br. s.,1H), 4.43 (d, J=10.3 Hz, 1H), 4.37 (d, J=10.0 Hz, 1H), 4.17 (q, J=7.2Hz, 2H), 2.79 (dd, J=17.8, 2.8 Hz, 1H), 2.46 (s, 3H), 2.43-2.17 (m, 4H),1.97-1.86 (m, 1H), 1.22 (t, J=7.3 Hz, 3H). LC/MS: m/e 430.2 (M+H)⁺, 2.20min.

Step 4. Preparation of ethyl1-(((5-methylisothiazol-3-yl)oxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 32% yield, following the proceduredescribed in general procedure A step 4, using ethyl1-(((5-methylisothiazol-3-yl)oxy)methyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-enecarboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 6.51-6.45 (m, 1H), 6.26(d, J=1.0 Hz, 1H), 4.42-4.37 (m, 1H), 4.33-4.26 (m, 1H), 4.11 (q, J=7.0Hz, 2H), 2.69-2.58 (m, 1H), 2.42 (d, J=1.0 Hz, 3H), 2.22-2.12 (m, 3H),2.02 (s, 1H), 1.95-1.87 (m, 1H), 1.82-1.74 (m, 1H), 1.22 (d, J=2.0 Hz,12H), 1.16 (t, J=7.2 Hz, 3H).

Step 5. Preparation of ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((5-methylisothiazol-3-yl)oxy)methyl)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 43% yield, following the proceduredescribed in general procedure A step 5, using ethyl1-(((5-methylisothiazol-3-yl)oxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-enecarboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 6.30 (d, J=1.0 Hz, 1H),5.33 (br. s., 1H), 5.17 (d, J=5.8 Hz, 1H), 4.70 (s, 2H), 4.58 (d, J=1.5Hz, 2H), 4.49-4.43 (m, 1H), 4.42-4.36 (m, 1H), 4.17-4.12 (m, 2H),3.13-2.96 (m, 8H), 2.73-2.62 (m, 2H), 2.62-2.52 (m, 2H), 2.50-2.41 (m,1H), 2.46 (d, J=1.0 Hz, 3H), 2.22-2.10 (m, 8H), 2.10-1.97 (m, 3H),1.96-1.65 (m, 4H), 1.68 (s, 3H), 1.64-1.37 (m, 7H), 1.37-1.23 (m, 6H),1.20 (t, J=7.1 Hz, 3H), 1.16-0.98 (m, 5H), 0.98-0.81 (m, 9H). LC/MS: m/e850.55 (M+H)⁺, 2.99 min (LCMS Method 3).

Step 6.4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((5-methylisothiazol-3-yl)oxy)methyl)cyclohex-3-ene-1-carboxylicacid was prepared in 36% yield, following the procedure described ingeneral procedure A step 6, using ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((5-methylisothiazol-3-yl)oxy)methyl)cyclohex-3-enecarboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 6.36 (d, J=0.8 Hz, 1H),5.37 (br. s., 1H), 5.30-5.10 (m, 1H), 4.79 (s, 1H), 4.72 (s, 1H), 4.51(d, J=10.0 Hz, 1H), 4.45 (dd, J=10.0, 3.5 Hz, 1H), 3.39 (d, J=12.3 Hz,1H), 3.28-2.87 (m, 11H), 2.86-2.57 (m, 2H), 2.49 (d, J=0.8 Hz, 3H),2.31-1.83 (m, 12H), 1.83-1.67 (m, 2H), 1.71 (s, 3H), 1.67-1.23 (m, 13H),1.16 (s, 3H), 1.13-1.02 (m, 2H), 1.06 (s, 3H), 0.97 (m, 3H), 0.93 (m,3H), 0.88 (s, 3H). LC/MS: m/e 822.60 (M+H)⁺, 2.83 min (LCMS Method 3).

Example 13 Preparation of4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((1-methyl-1H-tetrazol-5-yl)oxy)methyl)cyclohex-3-ene-1-carboxylicacid

Step 1. Preparation of ethyl8-(((1-methyl-1H-tetrazol-5-yl)oxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate

The title compound was prepared in 82% yield, following the proceduredescribed in general procedure A step 1-A, using1-methyl-1H-tetrazol-5-ol as reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ4.57 (s, 2H), 4.18 (q, J=7.0 Hz, 2H), 3.97-3.92 (m, 4H), 3.77 (s, 3H),2.29-2.21 (m, 1H), 2.18-2.10 (m, 1H), 1.76-1.63 (m, 6H), 1.24 (t, J=7.2Hz, 3H). LC/MS: m/e 327.20 (M+H)⁺, 2.15 min (LCMS Method 3).

Step 2. Preparation of ethyl1-(((1-methyl-1H-tetrazol-5-yl)oxy)methyl)-4-oxocyclohexane-1-carboxylate

The title compound was prepared in 91% yield, following the proceduredescribed in general procedure A step 2, using ethyl8-(((1-methyl-1H-tetrazol-5-yl)oxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 4.67 (s, 2H), 4.27 (q,J=7.2 Hz, 2H), 3.79 (s, 3H), 2.61-2.36 (m, 6H), 1.92-1.75 (m, 2H), 1.28(t, J=7.3 Hz, 3H). LC/MS: m/e 283.15 (M+H)⁺, 3.01 min (LCMS Method 10).

Step 3. Preparation of ethyl1-(((1-methyl-1H-tetrazol-5-yl)oxy)methyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 29% yield, following the proceduredescribed in general procedure A step 3, using ethyl1-(((1-methyl-1H-tetrazol-5-yl)oxy)methyl)-4-oxocyclohexanecarboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 5.80-5.72 (m, 1H),4.70-4.57 (m, 2H), 4.22-4.15 (m, 2H), 3.77 (s, 3H), 2.89-2.81 (m, 1H),2.50-2.23 (m, 4H), 1.97-1.88 (m, 1H), 1.25 (t, J=7.2 Hz, 3H). LC/MS: m/e415.25 (M+H)⁺, 2.51 min (LCMS Method 3).

Step 4. Preparation of ethyl1-(((1-methyl-1H-tetrazol-5-yl)oxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 90% yield, following the proceduredescribed in general procedure A step 4, using ethyl1-(((1-methyl-1H-tetrazol-5-yl)oxy)methyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-enecarboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 6.54-6.40 (m, 1H), 4.64(d, J=9.8 Hz, 1H), 4.56 (d, J=9.8 Hz, 1H), 4.17-4.10 (m, 2H), 3.74 (s,3H), 2.72-2.63 (m, 1H), 2.34-2.11 (m, 3H), 2.00-1.92 (m, 1H), 1.88-1.80(m, 1H), 1.23 (s, 12H), 1.21 (t, J=7.2 Hz 3H). LC/MS: m/e 393.35 (M+H)⁺,4.06 min (LCMS Method 10).

Step 5. Preparation of ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((1-methyl-1H-tetrazol-5-yl)oxy)methyl)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 56% yield, following the proceduredescribed in general procedure A step 5, using ethyl1-(((1-methyl-1H-tetrazol-5-yl)oxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-enecarboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 5.33 (br. s., 1H), 5.17(d, J=5.8 Hz, 1H), 4.70 (d, J=2.0 Hz, 1H), 4.58 (d, J=1.3 Hz, 1H),4.20-4.10 (m, 4H), 3.75 (s, 3H), 3.12-2.97 (m, 8H), 2.76-2.40 (m, 6H),2.26-0.87 (m, 27H), 1.68 (s, 3H), 1.21 (t, J=7.2 Hz, 3H), 1.04 (s, 3H),0.95 (s, 3H), 0.94-0.87 (m, 6H), 0.84 (s, 3H). LC/MS: m/e 835.60 (M+H)⁺,2.82 min (LCMS Method 3).

Step 6.4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((1-methyl-1H-tetrazol-5-yl)oxy)methyl)cyclohex-3-ene-1-carboxylicacid was prepared in 74% yield, following the procedure described ingeneral procedure A step 6, using ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((1-methyl-1H-tetrazol-5-yl)oxy)methyl)cyclohex-3-enecarboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 5.37 (br. s., 1H), 5.20(t, J=6.4 Hz, 1H), 4.79-4.61 (m, 4H), 3.79 (s, 2H), 3.26-2.98 (m, 10H),2.82 (d, J=9.3 Hz, 4H), 2.76-2.55 (m, 1H), 2.33-2.11 (m, 1H), 2.08 (s,3H), 2.11-1.82 (m, 8H), 1.70 (s, 3H), 1.65-1.37 (m, 10H), 1.36-1.22 (m,4H), 1.16 (s, 3H), 1.11-1.01 (m, 2H), 1.03 (s, 3H), 0.98 (s, 1.5H), 0.97(s, 1.5H), 0.94 (s, 1.5H), 0.93 (s, 1.5H), 0.87 (s, 3H). LC/MS: m/e807.60 (M+H)⁺, 2.90 min (LCMS Method 3).

Example 14 Preparation of4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((5-oxo-4,5-dihydro-1,2,4-thiadiazol-3-yl)oxy)methyl)cyclohex-3-ene-1-carboxylicacid

Step 1. Preparation of ethyl8-(((5-(methylthio)-1,2,4-thiadiazol-3-yl)oxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate

The title compound was prepared in 90% yield, following the proceduredescribed in general procedure A step 1-A, using5-(methylthio)-1,2,4-thiadiazol-3-ol as reactant. ¹H NMR (400 MHz,CHLOROFORM-d) δ 4.45 (s, 2H), 4.17 (q, J=7.0 Hz, 2H), 3.98-3.91 (m, 4H),2.68 (s, 3H), 2.31-2.21 (m, 2H), 1.77-1.66 (m, 6H), 1.24 (t, J=7.2 Hz,3H). LC/MS: m/e 375.10 (M+H)⁺, 2.50 min (LCMS Method 3).

Step 2. Preparation of ethyl1-(((5-(methylthio)-1,2,4-thiadiazol-3-yl)oxy)methyl)-4-oxocyclohexanecarboxylate

The title compound was prepared in 100% yield, following the proceduredescribed in general procedure A step 2, using ethyl8-(((5-(methylthio)-1,2,4-thiadiazol-3-yl)oxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 4.53 (s, 2H), 4.25 (q,J=7.1 Hz, 2H), 2.69 (s, 3H), 2.58-2.48 (m, 4H), 2.44-2.35 (m, 2H),1.96-1.86 (m, 2H), 1.28 (t, J=12 Hz, 3H). LC/MS: m/e 331.05 (M+H)⁺, 2.32min (LCMS Method 3).

Step 3. Preparation of ethyl1-(((5-(methylthio)-1,2,4-thiadiazol-3-yl)oxy)methyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 55% yield, following the proceduredescribed in general procedure A step 3, using ethyl1-(((5-(methylthio)-1,2,4-thiadiazol-3-yl)oxy)methyl)-4-oxocyclohexanecarboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 5.74 (td, J=3.1, 1.8 Hz,1H), 4.50 (d, J=10.3 Hz, 1H), 4.45 (d, J=10.3 Hz, 1H), 4.15 (q, J=7.0Hz, 2H), 2.84-2.75 (m, 1H), 2.65 (s, 3H), 2.51-2.19 (m, 4H), 2.02-1.94(m, 1H), 1.21 (t, J=7.2 Hz, 3H).

Step 4. Preparation of ethyl1-(((5-(methylthio)-1,2,4-thiadiazol-3-yl)oxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-enecarboxylate

The title compound was prepared in 39% yield, following the proceduredescribed in general procedure A step 4 for 7 h, using ethyl1-(((5-(methylthio)-1,2,4-thiadiazol-3-yl)oxy)methyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-enecarboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 6.50 (dt, J=3.5, 1.8 Hz,1H), 4.49 (d, J=10.0 Hz, 1H), 4.40 (d, J=10.0 Hz, 1H), 4.11 (q, J=7.0Hz, 2H), 2.72-2.64 (m, 1H), 2.64 (s, 3H), 2.28-2.16 (m, 3H), 1.98-1.81(m, 2H), 1.23 (s, 12H), 1.17 (t, J=7.2 Hz, 3H). LC/MS: m/e 441.25(M+H)⁺, 2.92 min (LCMS Method 3).

Step 5. Preparation of ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((5-(methylthio)-1,2,4-thiadiazol-3-yl)oxy)methyl)cyclohex-3-enecarboxylate

The title compound was prepared, following the procedure described ingeneral procedure A step 5 at 90° C. for 4 h, using ethyl1-(((5-(methylthio)-1,2,4-thiadiazol-3-yl)oxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-enecarboxylateas reactant. LC/MS: m/e 883.55 (M+H)⁺, 3.11 min (LCMS Method 3).

Step 6.4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((5-oxo-4,5-dihydro-1,2,4-thiadiazol-3-yl)oxy)methyl)cyclohex-3-ene-1-carboxylicacid was prepared in 10% yield, following the procedure described ingeneral procedure A step 6, using ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((5-(methylthio)-1,2,4-thiadiazol-3-yl)oxy)methyl)cyclohex-3-enecarboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 4.80 (s, 1H), 4.76-4.69(m, 1H), 4.60-4.50 (m, 2H), 3.35-3.01 (m, 10H), 3.01-2.78 (m, 4H),2.67-2.51 (m, 4H), 2.51-2.36 (m, 5H), 2.36-2.14 (m, 2H), 1.81-1.75 (m,2H), 1.72 (s, 3H), 1.75-1.68 (m, 2H), 1.68-1.34 (m, 11H), 1.27 (s, 3H),1.26 (s, 3H), 1.20 (s, 3H), 1.08 (s, 3H), 1.13-1.03 (m, 4H), 0.94 (s,3H). LC/MS: m/e 825.50 (M+H)⁺, 2.78 min (LCMS Method 3).

Example 15 Preparation of4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((thiazol-2-yloxy)methyl)cyclohex-3-ene-1-carboxylicacid

Step 1: Preparation of ethyl8-((thiazol-2-yloxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate

The title compound was prepared in 35% yield, following the proceduredescribed in general procedure A step 1-A, using thiazol-2-ol asreactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 6.39 (d, J=5.5 Hz, 1H), 6.03(d, J=5.3 Hz, 1H), 4.09 (q, J=7.3 Hz, 2H), 3.86 (s, 4H), 3.76 (s, 2H),2.11-2.03 (m, 2H), 1.68-1.46 (m, 6H), 1.20 (t, J=7.2 Hz, 3H). LC/MS m/z328.10 (M+H)⁺, 2.09 min (LCMS Method 3).

Step 2. Preparation of ethyl4-oxo-1-((thiazol-2-yloxy)methyl)cyclohexane-1-carboxylate

The title compound was prepared in 80% yield, following the proceduredescribed in general procedure A step 2, using ethyl8-((thiazol-2-yloxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate asreactant. ¹H NMR (400 MHz, CHLOROFORM-d) □ 6.47 (d, J=5.5 Hz, 1H), 6.13(d, J=5.5 Hz, 1H), 4.28 (q, J=7.0 Hz, 2H), 3.97 (s, 2H), 2.55-2.35 (m,6H), 1.89-1.77 (m, 2H), 1.34 (t, J=7.2 Hz, 3H). MS m/z 284.20 (M+H)⁺,1.72 min (LCMS Method 3).

Step 3. Preparation of ethyl1-((thiazol-2-yloxy)methyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 22% yield as an oil, following theprocedure described in general procedure A step 3, using ethyl4-oxo-1-((thiazol-2-yloxy)methyl)cyclohexane-1-carboxylate as reactant.¹H NMR (400 MHz, CHLOROFORM-d) δ 6.43 (d, J=5.5 Hz, 1H), 6.11 (d, J=5.3Hz, 1H), 5.73 (td, J=3.4, 1.5 Hz, 1H), 4.16 (qd, J=7.2, 2.6 Hz, 2H),3.90 (s, 2H), 2.74-2.64 (m, 1H), 2.45-2.39 (m, 2H), 2.33-2.20 (m, 2H),1.85-1.76 (m, 1H), 1.25 (t, J=7.2 Hz, 3H). MS m/z 416.20 (M+H)⁺, 2.75min (LCMS Method 3).

Step 4. Preparation of ethyl4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-((thiazol-2-yloxy)methyl)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 71% yield, following the proceduredescribed in general procedure A step 4, using ethyl1-((thiazol-2-yloxy)methyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-enecarboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 6.49-6.46 (m, 1H), 6.45(d, J=5.3 Hz, 1H), 6.06 (d, J=5.3 Hz, 1H), 4.12 (qd, J=7.2, 2.6 Hz, 2H),3.86 (s, 2H), 2.63-2.54 (m, 1H), 2.31-1.99 (m, 4H), 1.60 (ddd, J=13.0,9.0, 5.6 Hz, 1H), 1.23 (s, 12H), 1.22 (t, J=7.2 Hz, 3H). MS m/z 394.30(M+H)⁺, 2.65 min (LCMS Method 3).

Step 5. Preparation of ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((thiazol-2-yloxy)methyl)cyclohex-3-enecarboxylate

The title compound was prepared in 30% yield as a solid, following theprocedure described in general procedure A step 5, using(1R,3aS,5aR,5bR,7aR,11aR,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yltrifluoromethanesulfonate and ethyl4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-((thiazol-2-yloxy)methyl)cyclohex-3-enecarboxylateas reactants. NMR (400 MHz, CHLOROFORM-d) δ 6.46 (d, J=5.5 Hz, 1H), 6.06(d, J=5.3 Hz, 1H), 5.30 (br. s., 1H), 5.18-5.13 (m, 1H), 4.70 (d, J=2.0Hz, 1H), 4.58 (s, 1H), 4.11 (q, J=7.3 Hz, 2H), 3.96-3.84 (m, 2H),3.11-2.97 (m, 8H), 2.74-2.42 (m, 6H), 2.22-0.85 (m, 27H), 1.67 (s, 3H),1.25 (t, J=7.2 Hz, 3H), 1.04 (s, 3H), 0.94 (s, 3H), 0.93-0.86 (m, 6H),0.83 (s, 3H). MS m/z 836.65 (M+H)⁺, 2.98 min (LCMS Method 3).

Step 6.4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((thiazol-2-yloxy)methyl)cyclohex-3-ene-1-carboxylicacid was prepared in 68% yield as a solid, following the proceduredescribed in general procedure A step 6, using ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((thiazol-2-yloxy)methyl)cyclohex-3-enecarboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 6.70 (d, J=5.5 Hz, 1H),6.08 (d, J=5.5 Hz, 1H), 5.42-5.28 (m, 1H), 5.20 (dd, J=16.2, 4.9 Hz,1H), 4.78 (s, 1H), 4.69 (s, 1H), 4.19-4.01 (m, 1H), 4.02-3.85 (m, 1H),3.29 (d, J=15.8 Hz, 1H), 3.24-2.95 (m, 7H), 2.85 (d, J=10.8 Hz, 2H),2.61 (d, J=16.6 Hz, 1H), 2.43 (d, J=15.1 Hz, 1H), 2.31-2.12 (m, 8H),2.12-1.85 (m, 6H), 1.85-1.75 (m, 1H), 1.70 (s, 3H), 1.75-1.60 (m, 2H),1.59-1.21 (m, 12H), 1.17 (s, 3H), 1.13-1.01 (m, 2H), 1.04 (s, 3H), 0.98(s, 3H), 0.96 (s, 3H), 0.86 (s, 3H). LC/MS: m/e 808.55 (M+H)⁺, 1.832 min(LCMS Method 3).

Example 16 Preparation of4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((1-phenyl-1H-1,2,3-triazol-5-yl)oxy)methyl)cyclohex-3-ene-1-carboxylicacid

Step 1. Preparation of ethyl8-(((1-phenyl-1H-1,2,3-triazol-5-yl)oxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate

The title compound was prepared following the procedure described ingeneral procedure A step 1-A, using 1-phenyl-1H-1,2,3-triazol-5-ol asreactant. This material was carried forward to the next step withoutpurification. LC/MS: m/e 388.20 (M+H)⁺, 2.32 min (LCMS Method 3).

Step 2. Preparation of ethyl4-oxo-1-(((1-phenyl-1H-1,2,3-triazol-5-yl)oxy)methyl)cyclohexanecarboxylate

The title compound was prepared in 9% yield, following the proceduredescribed in general procedure A step 2, using ethyl8-(((1-phenyl-1H-1,2,3-triazol-5-yl)oxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.66-7.59 (m, 2H),7.53-7.45 (m, 1H), 7.45-7.36 (m, 2H), 7.20 (s, 1H), 4.22 (s, 2H), 4.15(q, J=7.1 Hz, 2H), 2.53-2.42 (m, 4H), 2.40-2.30 (m, 2H), 1.83-1.71 (m,2H), 1.15 (t, J=7.2 Hz, 3H). LC/MS: m/e 388.20 (M+H)⁺, 2.32 min (LCMSMethod 3).

Step 3. Preparation of ethyl1-(((1-phenyl-1H-1,2,3-triazol-5-yl)oxy)methyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-enecarboxylate

The title compound was prepared in 144% yield, following the proceduredescribed in general procedure A step 3, using ethyl4-oxo-1-(((1-phenyl-1H-1,2,3-triazol-5-yl)oxy)methyl)cyclohexanecarboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.69-7.63 (m, 2H),7.54-7.48 (m, 2H), 7.46-7.40 (m, 1H), 7.22 (s, 1H), 5.80-5.75 (m, 1H),4.29 (d, J=9.0 Hz, 1H), 4.22 (d, J=8.8 Hz, 1H), 4.17-4.11 (m, 2H),2.87-2.79 (m, 1H), 2.56-2.44 (m, 1H), 2.42-2.22 (m, 3H), 1.97-1.89 (m,1H), 1.17 (t, J=7.2 Hz, 3H). LC/MS: m/e 476.25 (M+H)⁺, 2.65 min (LCMSMethod 3).

Step 4. Preparation of ethyl1-(((1-phenyl-1H-1,2,3-triazol-5-yl)oxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-enecarboxylate

The title compound was prepared in 91% yield as an oil, following theprocedure described in general procedure A step 4, using ethyl1-(((1-phenyl-1H-1,2,3-triazol-5-yl)oxy)methyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-enecarboxylateas reactant.

¹H NMR (400 MHz, CHLOROFORM-d) δ 7.68-7.63 (m, 2H), 7.50-7.43 (m, 2H),7.41-7.35 (m, 1H), 7.19 (s, 1H), 6.49 (dt, J=3.5, 1.8 Hz, 1H), 4.25 (d,J=8.8 Hz, 1H), 4.19 (d, J=8.8 Hz, 1H), 4.08 (qd, J=7.1, 1.0 Hz, 2H),2.69-2.60 (m, 1H), 2.28-2.05 (m, 3H), 1.98-1.90 (m, 1H), 1.88-1.81 (m,1H), 1.23 (s, 12H), 1.12 (t, J=7.2 Hz, 3H). LC/MS: m/e 454.35 (M+H)⁺,2.63 min (LCMS Method 3).

Step 4. Preparation of ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((1-phenyl-1H-1,2,3-triazol-5-yl)oxy)methyl)cyclohex-3-enecarboxylate

4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((1-phenyl-1H-1,2,3-triazol-5-yl)oxy)methyl)cyclohex-3-ene-1-carboxylicacid was prepared in 58% yield as a solid, following the proceduredescribed in general procedure A step 5, using ethyl1-(((1-phenyl-1H-1,2,3-triazol-5-yl)oxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-enecarboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.71-7.65 (m, 2H),7.51-7.44 (m, 2H), 7.41-7.35 (m, 1H), 7.19 (s, 1H), 5.32 (br. s., 1H),5.15 (d, J=4.8 Hz, 1H), 4.68 (d, J=2.0 Hz, 1H), 4.57 (s, 1H), 4.30-4.20(m, 2H), 4.09 (q, J=7.3 Hz, 2H), 3.09-2.96 (m, 8H), 2.71-2.38 (m, 6H),2.25-0.86 (m, 27H), 1.66 (s, 3H), 1.14 (t, J=7.2 Hz, 3H), 1.03 (s, 3H),0.94 (s, 3H), 0.93-0.87 (m, 6H), 0.83 (s, 3H).

Step 6. The title compound was prepared in 20% yield as a solid,following the procedure described in general procedure A step 5, usingethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((1-phenyl-1H-1,2,3-triazol-5-yl)oxy)methyl)cyclohex-3-enecarboxylate as reactant. ¹H NMR (400MHz, CHLOROFORM-d) δ 7.79-7.70 (m, 2H), 7.53-7.46 (m, 2H), 7.43-7.37 (m,1H), 7.26-7.22 (m, 1H), 5.36 (br. s., 1H), 5.18 (t, J=5.5 Hz, 1H), 4.70(s, 1H), 4.61 (s, 1H), 4.38-4.23 (m, 2H), 3.09-2.92 (m, 8H), 2.90-2.80(m, 2H), 2.78-2.54 (m, 4H), 2.31-2.10 (m, 4H), 2.04-1.80 (m, 6H), 1.73(d, J=11.3 Hz, 1H), 1.67 (s, 3H), 1.54 (d, J=17.8 Hz, 3H), 1.49-1.35 (m,6H), 1.35-1.15 (m, 5H), 1.11 (s, 3H), 1.14-1.02 (m, 2H), 1.00 (s, 3H),0.97-0.94 (m, 1H), 0.96 (s, 3H), 0.93-0.92 (m, 3H), 0.85 (s, 3H). LC/MS:m/e 868.65 (M+H)⁺, 2.83 min (LCMS Method 3).

Example 17 Preparation of4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((1-isopropyl-1H-1,2,3-triazol-5-yl)oxy)methyl)cyclohex-3-ene-1-carboxylicacid

Step 1. Preparation of ethyl8-(((1-isopropyl-1H-1,2,3-triazol-5-yl)oxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate

The title compound was prepared in 43% yield as a semi-solid, followingthe procedure described in general procedure A step 1-B at 105° C.,using 1-isopropyl-1H-1,2,3-triazol-5-ol as reactant. ¹H NMR (400 MHz,CHLOROFORM-d) δ 7.06 (s, 1H), 4.59 (spt, J=6.8 Hz, 1H), 4.18 (q, J=7.2Hz, 2H), 4.08 (s, 2H), 4.00-3.91 (m, 4H), 2.33-2.24 (m, 2H), 1.76-1.65(m, 6H), 1.51 (d, J=6.8 Hz, 6H), 1.24 (t, J=7.2 Hz, 3H). LC/MS: m/e354.30 (M+H)⁺, 3.33 min (LCMS Method 11).

Step 2. Preparation of ethyl1-(((1-isopropyl-1H-1,2,3-triazol-5-yl)oxy)methyl)-4-oxocyclohexane-1-carboxylate

The title compound was prepared in 91% yield as an oil, following theprocedure described in general procedure A step 2, using ethyl8-(((1-isopropyl-1H-1,2,3-triazol-5-yl)oxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.02 (s, 1H), 4.50 (spt,J=6.8 Hz, 1H), 4.17 (q, J=7.0 Hz, 2H), 4.11 (s, 2H), 2.52-2.40 (m, 4H),2.38-2.28 (m, 2H), 1.82-1.71 (m, 2H), 1.43 (d, J=7.0 Hz, 6H), 1.19 (t,J=1.2 Hz, 3H). LC/MS: m/e 354.30 (M+H)⁺, 1.96 min (LCMS Method 3).

Step 3. Preparation of ethyl1-(((1-isopropyl-1H-1,2,3-triazol-5-yl)oxy)methyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-enecarboxylate

The title compound was prepared in 97% yield as an oil, following theprocedure described in general procedure A step 3, using ethyl1-(((1-isopropyl-1H-1,2,3-triazol-5-yl)oxy)methyl)-4-oxocyclohexanecarboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.04 (s, 1H), 5.79-5.73(m, 1H), 4.53 (spt, J=6.8 Hz, 1H), 4.19-4.04 (m, 4H), 2.88-2.76 (m, 1H),2.55-2.21 (m, 4H), 1.92 (ddd, J=13.7, 7.9, 6.3 Hz, 1H), 1.47 (d, J=6.8Hz, 6H), 1.20 (t, J=7.2 Hz, 3H). ¹⁹F NMR (376 MHz, CHLOROFORM-d) □-73.94(s, 3F). LC/MS: m/e 442.20 (M+H)⁺, 2.64 min (LCMS Method 3).

Step 4. Preparation of ethyl1-(((1-isopropyl-1H-1,2,3-triazol-5-yl)oxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-enecarboxylate

The title compound was prepared in 100% yield, following the proceduredescribed in general procedure A step 4, using ethyl1-(((1-isopropyl-1H-1,2,3-triazol-5-yl)oxy)methyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-enecarboxylateas reactant.

¹H NMR (400 MHz, CHLOROFORM-d) δ 7.02 (s, 1H), 6.48 (dt, J=3.3, 1.7 Hz,1H), 4.53 (spt, J=6.8 Hz, 1H), 4.16-4.06 (m, 4H), 2.69-2.60 (m, 1H),2.28-2.05 (m, 3H), 1.98-1.81 (m, 2H), 1.46 (dd, J=6.8, 2.3 Hz, 6H), 1.22(s, 12H), 1.17 (t, J=1.2 Hz, 3H). LC/MS: m/e 420.30 (M+H)⁺, 2.65 min(LCMS Method 3).

Step 5. Preparation of ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((1-isopropyl-1H-1,2,3-triazol-5-yl)oxy)methyl)cyclohex-3-enecarboxylate

The title compound was prepared in 100% yield, following the proceduredescribed in general procedure A step 5, using ethyl1-(((1-isopropyl-1H-1,2,3-triazol-5-yl)oxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-enecarboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.05 (s, 1H), 5.33 (br.s., 1H), 5.16 (d, J=4.8 Hz, 1H), 4.68 (s, 1H), 4.56 (s, 1H), 4.56 (spt,J=6.7 Hz, 1H), 4.20-4.09 (m, 4H), 3.11-2.93 (m, 8H), 2.71-2.36 (m, 6H),2.30-0.86 (m, 27H), 1.66 (s, 3H), 1.49 (d, J=6.3 Hz, 6H), 1.21 (t, J=7.2Hz, 3H), 1.04 (s, 3H), 0.94 (s, 3H), 0.93-0.87 (m, 6H), 0.83 (s, 3H).LC/MS: m/e 862.73 (M+H)⁺, 2.35 min (LCMS Method 1).

Step 6.4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((1-isopropyl-1H-1,2,3-triazol-5-yl)oxy)methyl)cyclohex-3-ene-1-carboxylicaad was prepared in 45% yield, following the procedure described ingeneral procedure A step 6, using ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((1-isopropyl-1H-1,2,3-triazol-5-yl)oxy)methyl)cyclohex-3-enecarboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.12 (d, J=2.3 Hz, 1H),5.36 (br. s., 1H), 5.19 (d, J=4.3 Hz, 1H), 4.72 (s, 1H), 4.63 (s, 1H),4.67-4.52 (h, J=6.8 Hz, 1H), 4.32-4.10 (m, 2H), 3.20-2.89 (m, 8H),2.87-2.68 (m, 3H), 2.68-2.53 (m, 1H), 2.34-2.21 (m, 1H), 2.21-1.85 (m,11H), 1.85-1.73 (m, 1H), 1.71-1.65 (m. 1H), 1.68 (s, 3H), 1.51 (d, J=6.5Hz, 6H), 1.64-1.36 (m, 9H), 1.36-1.19 (m, 4H), 1.14 (s, 3H), 1.07 (br.s., 2H), 1.01 (s, 3H), 0.97-0.96 (m, 4H), 0.94-0.89 (m, 3H), 0.87 (s,3H). LC/MS: m/e 834.69 (M+H)⁺, 2.32 min (LCMS Method 1).

Example 18 Preparation of4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((5-(prop-1-en-2-yl)isothiazol-3-yl)oxy)methyl)cyclohex-3-ene-1-carboxylicacid

Step 1. Preparation of ethyl8-((isothiazol-3-yloxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate

The title compound was prepared in 36% yield, following the proceduredescribed in general procedure A step 1-A, using isothiazol-3(2H)-one asreactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 8.42 (d, J=4.8 Hz, 1H), 6.57(d, J=4.8 Hz, 1H), 4.42 (s, 2H), 4.17 (q, J=7.0 Hz, 2H), 3.99-3.93 (m,4H), 2.31-2.20 (m, 2H), 1.75-1.65 (m, 6H), 1.23 (t, J=7.2 Hz, 3H).LC/MS: m/e 328.20 (M+H) 3.59 min (LCMS Method 12).

Step 2. Preparation of ethyl8-(((5-(2-hydroxypropan-2-yl)isothiazol-3-yl)oxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate

To a solution of ethyl8-((isothiazol-3-yloxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate(100 mg, 0305 mmol) in THF (2 mL) under nitrogen at −78° C. was added a2M solution of LDA (0.305 mL, 0.611 mmol). It was stirred at −78° C. for20 minutes before it was added neat propan-2-one (0.045 mL, 0.611 mmol).Stirring continued for another 30 minutes at −78° C. The reaction wasquenched with a half-saturated aminonium chloride in 0.5M HCl, extractedwith ethyl acetate and concentrated in vacuo. The crude mixture waspurified by silica gel column eluted with 0-45% EtOAc/hexanes to givethe desired product as an oil (83 mg, 70%). ¹H NMR (400 MHz,CHLOROFORM-d) δ 6.35 (s, 1H), 4.32 (s, 2H), 4.13 (q, J=7.1 Hz, 2H), 3.91(s, 4H), 2.84 (s, 1H), 2.26-2.12 (m, 2H), 1.72-1.61 (m, 6H), 1.58 (s,6H), 1.19 (t, J=7.2 Hz, 3H). LC/MS: m/e 386.20 (M+H)⁺, 2.75 min (LCMSMethod 13).

Step 3. Preparation of ethyl1-(((5-(2-hydroxypropan-2-yl)isothiazol-3-yl)oxy)methyl)-4-oxocyclohexanecarboxylate

The title compound was prepared in 100% yield, following the proceduredescribed in general procedure A step 2 using ethyl8-(((5-(2-hydroxypropan-2-yl)isothiazol-3-yl)oxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 6.36 (s, 1H), 4.41 (s,2H), 4.20 (q, J=7.2 Hz, 2H), 2.99 (s, 1H), 2.52-2.41 (m, 4H), 2.39-2.29(m, 2H), 1.88-1.75 (m, 2H), 1.59 (s, 6H), 1.23 (t, J=7.0 Hz, 3H). LC/MS:m/e 342.15 (M+H)⁺, 2.03 min (LCMS Method 3).

Step 4. Preparation of ethyl1-(((5-(prop-1-en-2-yl)isothiazol-3-yl)oxy)methyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 22% yield, following the proceduredescribed in general procedure A step 3 using ethyl1-(((5-(2-hydroxypropan-2-yl)isothiazol-3-yl)oxy)methyl)-4-oxocyclohexanecarboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 6.50 (s, 1H), 5.76 (td,J=3.3, 1.8 Hz, 1H), 5.45 (s, 1H), 5.18 (s, 1H), 4.47 (d, J=10.0 Hz, 1H),4.41 (d, J=10.0 Hz, 1H), 4.19 (qd, J=7.1, 0.8 Hz, 2H), 2.85-2.77 (m,1H), 2.53-2.22 (m, 4H), 2.09 (s, 3H), 1.98-1.90 (m, 1H), 1.24 (t, J=7.2Hz, 3H). LC/MS: m/e 456.10 (M+H)⁺, 2.76 min (LCMS Method 3).

Step 5. Preparation of ethyl1-(((5-(prop-1-en-2-yl)isothiazol-3-yl)oxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 78% yield, following the proceduredescribed in general procedure A step 4 using ethyl1-(((5-(prop-1-en-2-yl)isothiazol-3-yl)oxy)methyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-enecarboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 6.55-6.50 (m, 1H), 6.49(s, 1H), 5.43 (s, 1H), 5.14 (s, 1H), 4.46 (d, J=10.0 Hz, 1H), 4.37 (d,J=10.0 Hz, 1H), 4.15 (q, J=7.0 Hz, 2H), 2.74-2.64 (m, 1H), 2.28-2.16 (m,3H), 2.09 (s, 3H), 2.01-1.80 (m, 2H), 1.26 (s, 12H), 1.21 (t, J=7.2 Hz,3H). LC/MS: m/e 434.20 (M+H)⁺, 2.79 min (LCMS Method 3).

Step 6. Preparation of ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((5-(prop-1-en-2-yl)isothiazol-3-yl)oxy)methyl)cyclohex-3-enecarboxylate

The title compound was prepared in 42% yield, following the proceduredescribed in general procedure A step 5 using ethyl1-(((5-(prop-1-en-2-yl)isothiazol-3-yl)oxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-enecarboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 6.51 (s, 1H), 5.44 (s,1H), 5.35 (br. s., 1H), 5.19 (br. s., 1H), 5.17 (s, 1H), 4.76 (s, 1H),4.71 (s, 1H), 4.51-4.38 (m, 2H), 4.21-4.12 (m, 2H), 3.41-2.92 (m, 11H),2.78-2.54 (m, 3H), 2.22-0.89 (m, 27H), 2.09 (s, 3H), 1.69 (s, 3H), 1.23(t, J=7.2 Hz, 3H), 1.13 (s, 3H), 1.04 (s, 3H), 0.96-0.91 (m, 6H), 0.87(s, 3H). LC/MS: m/e 876.60 (M+H)⁺, 3.01 min (LCMS Method 3).

Step 7.4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((5-(prop-1-en-2-yl)isothiazol-3-yl)oxy)methyl)cyclohex-3-ene-1-carboxylicacid was prepared in 56% yield, following the procedure described ingeneral procedure A step 6 using ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((5-(prop-1-en-2-yl)isothiazol-3-yl)oxy)methyl)cyclohex-3-enecarboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 6.55 (s, 1H), 5.47 (s,1H), 5.37 (br. s., 1H), 5.21 (d, J=5.3 Hz, 1H), 5.19 (s, 1H), 4.79 (s,1H), 4.73 (s, 1H), 4.56-4.50 (m, 1H), 4.50-4.43 (m, 1H), 3.40 (d, J=1.8Hz, 1H), 3.29-2.91 (m, 10H), 2.80-2.72 (m, 1H), 2.72-2.62 (m, 1H),2.34-2.09 (m, 6H), 2.11 (s, 3H), 2.09-1.97 (m, 4H), 1.97-1.83 (m, 2H),1.83-1.68 (m, 2H), 1.71 (s, 3H), 1.67-1.37 (m, 12H), 1.37-1.23 (m, 1H),1.15 (s, 3H), 1.13-1.03 (m, 2H), 1.06 (s, 3H), 0.98-0.97 (m, 3H),0.95-0.93 (m, 3H), 0.89 (s, 3H). LC/MS: m/e 848.50 (M+H)⁺, 3.05 min(LCMS Method 3).

Example 19 Preparation of4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridazin-3-yloxy)methyl)cyclohex-3-ene-1-carboxylicacid

Step 1. Preparation of ethyl8-((pyridazin-3-yloxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate

To the solution of ethyl8-(hydroxymethyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate (300 mg, 1.23mmol) in DMF (6 mL) at 0° C. was added potassium tert-butoxide (1.84 mL,1.84 mmol) followed by 3-chloropyridazine (211 mg, 1.84 mmol). Theresulting suspension was stirred at 0° C. then warmed to RT overnight.The reaction mixture was diluted with ethyl acetate (10 mL), washed withwater, dried over sodium sulfate, and concentrated in vacuo to givecrude product. LC/MS: m/e 323.20 (M+H)⁺, 2.09 min (LCMS Method 7).

Step 2. Preparation of ethyl4-oxo-1-((pyridazin-3-yloxy)methyl)cyclohexanecarboxylate

The title compound was prepared in 70% yield, following the proceduredescribed in general procedure A step 2 using ethyl8-((pyridazin-3-yloxy)methyl)-1,4-dioxaspiro[4.5]decane-8-carboxylate asreactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 8.82 (dd, J=4.5, 1.3 Hz, 1H),7.37 (dd, J=9.0, 4.5 Hz, 1H), 6.96 (dd, J=8.9, MHz, 1H), 4.64 (s, 2H),4.21 (q, J=7.1 Hz, 2H), 2.57-2.28 (m, 6H), 1.92-1.82 (m, 2H), 1.28 (t,J=7.2 Hz, 3H). LC/MS: m/e 279.15 (M+H)⁺, 1.71 min (LCMS Method 7).

Step 3. Preparation of ethyl1-((pyridazin-3-yloxy)methyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-enecarboxylate

The title compound was prepared in 39% yield, following the proceduredescribed in general procedure A step 3 using ethyl4-oxo-1-((pyridazin-3-yloxy)methyl)cyclohexanecarboxylate as reactant.¹H NMR (400 MHz, CHLOROFORM-d) δ 8.82 (dd, J=4.5, MHz, 1H), 7.36 (dd,J=9.0, 4.5 Hz, 1H), 6.94 (dd, J=9.0, 1.3 Hz, 1H), 5.75 (td, J=3.1, 1.8Hz, 1H), 4.62 (d, J=10.5 Hz, 1H), 4.59 (d, J=10.5 Hz, 1H), 4.18-4.11 (m,2H), 2.88-2.79 (m, 1H), 2.53-2.23 (m, 4H), 1.97-1.90 (m, 1H), 1.21 (t,J=7.2 Hz, 3H). LC/MS: m/e 411.15 (M+H)⁺, 2.66 min (LCMS Method 7).

Step 4. Preparation of ethyl1-((pyridazin-3-yloxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-enecarboxylate

The title compound was prepared in 43% yield, following the proceduredescribed in general procedure A step 4 using ethyl1-((pyridazin-3-yloxy)methyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-enecarboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 8.78 (dd, J=4.5, 1.3 Hz,1H), 7.32 (dd, J=8.9, 4.4 Hz, 1H), 6.91 (dd, J=8.9, 1.4 Hz, 1H), 6.49(dt, J=3.7, 1.8 Hz, 1H), 4.62 (d, J=10.3 Hz, 1H), 4.53 (d, J=10.5 Hz,1H), 4.07 (q, J=7.0 Hz, 2H), 2.72-2.64 (m, 1H), 2.27-2.08 (m, 3H),1.98-1.80 (m, 2H), 1.21 (s, 12H), 1.21 (t, J=7.3 Hz, 3H). LC/MS: m/e389.25 (M+H)⁺, 2.74 min (LCMS Method 7).

Step 5. Preparation of ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridazin-3-yloxy)methyl)cyclohex-3-enecarboxylate

The title compound was prepared following the procedure described ingeneral procedure A step 5 using ethyl1-((pyridazin-3-yloxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-enecarboxylateas reactant. The crude material was taken directly into the next stepwithout purification.

Step 6.4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridazin-3-yloxy)methyl)cyclohex-3-ene-1-carboxylicacid was prepared in 22% yield as a solid, following the proceduredescribed in general procedure A step 6 using ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridazin-3-yloxy)methyl)cyclohex-3-enecarboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 9.09 (d, J=4.5 Hz, 1H),7.74 (dd, J=9.0, 4.5 Hz, 1H), 7.35 (d, J=8.8 Hz, 1H), 5.38 (br. s., 1H),5.21 (t, J=5.6 Hz, 1H), 4.79 (s, 1H), 4.72 (s, 1H), 4.76-4.64 (m, 2H),3.39 (d, J=12.5 Hz, 1H), 3.25-3.02 (m, 9H), 3.02-2.86 (m, 2H), 2.86-2.62(m, 2H), 2.32-2.06 (m, 5H), 2.06-1.84 (m, 6H), 1.82-1.67 (m, 2H), 1.71(s, 3H), 1.66-1.35 (m, 10H), 1.35-1.20 (m, 4H), 1.17 (s, 3H), 1.14-1.04(m, 2H), 1.05 (s, 3H), 0.97-0.95 (m, 3H), 0.92-0.91 (m, 3H), 0.87 (s,3H). LC/MS: m/e 803.48 (M+H)⁺, 2.27 min (LCMS Method 1).

General Procedure B: Preparation of (R) α-SubstitutedCyclohexenecarboxylic Acid Derivatives.

Step 1. Preparation of((R)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(ethoxycarbonyl)cyclohex-3-en-1-yl)methylbenzoate

A mixture of(1R,3aS,5aR,5bR,7aR,11aR,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yltrifluoromethanesulfonate (1 eq), (R)-(1-(ethoxycarbonyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-en-1-yl)methylbenzoate (1.2 eq), Na₂CO₃ (3 eq) and Pd(Ph₃P)₄ (0.06 eq) in 1,4-dioxaneand H₂O (4:1) was flushed with nitrogen, sealed and heated at 70° C. for2 h. The reaction mixture was diluted with EtOAc, washed with brine,dried over Na₂SO₄, and concentrated in vacuo. The crude product waspurified by silica gel column eluted with 0-35% Ethyl acetate/hexanes togive the desired product (68% yield) as a solid. ¹H NMR (400 MHz,CHLOROFORM-d) δ 8.01 (dd, J=8.4, 1.4 Hz, 2H), 7.60-7.53 (m, 1H),7.47-7.40 (m, 2H), 5.36 (br. s., 1H), 5.20 (dd, J=6.0, 1.8 Hz, 1H), 4.71(d, J=2.0 Hz, 1H), 4.60 (s, 1H), 4.49-4.39 (m, 2H), 4.18 (qd, J=7.2, 1.4Hz, 2H), 3.13-2.98 (m, 8H), 2.73-2.43 (m, 6H), 2.27-0.89 (m, 27H), 1.69(s, 3H), 1.25-1.20 (m, 3H), 1.07 (s, 3H), 0.97 (br. s., 3H), 0.96 (br.s., 3H), 0.94 (s, 3H), 0.87 (s, 3H). LC/MS m/z 857.65 (M+H)⁺, 2.43 mm(LCMS Method 1).

Step 2. Preparation of ethyl(R)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(hydroxymethyl)cyclohex-3-ene-1-carboxylate

A suspension of((R)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(ethoxycarbonyl)cyclohex-3-en-1-yl)methyl benzoate (1 eq) and 1N NaOH (1 eq) inMeOH and THF was stirred at RT for 2 days. The mixture was neutralizedwith 1N HCl and the solvent was removed in vacuo. The residue was takeninto CH₂Cl₂, washed with H₂O followed by brine, dried over Na₂SO₄, andconcentrated in vacuo. The crude product was purified on silica geleluted with ethyl acetate/hexanes to give the desired product (85%yield) as a solid. ¹H NMR (400 MHz, CHLOROFORM-d) δ 5.32 (br. s., 1H),5.18 (d, J=4.8 Hz, 1H), 4.71 (d, J=2.0 Hz, 1H), 4.60 (s, 1H), 4.19 (q,J=7.2 Hz, 2H), 3.69 (br. s., 2H), 3.12-2.98 (m, 8H), 2.72-2.43 (m, 6H),2.28-0.89 (m, 27H), 1.70 (s, 3H), 1.28 (t, J=7.2 Hz, 3H), 1.07 (s, 3H),0.97 (s, 3H), 0.96 (s, 3H), 0.93 (s, 3H), 0.86 (s, 3H). LC/MS m/z 753.65(M+H)⁺, 3.79 min (LCMS Method 2).

Step 3. Preparation of (R) α-methyl ether

To a solution of ethyl(R)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(hydroxymethyl)cyclohex-3-ene-1-carboxylate(1 eq) and Ar—X (2 eq) in DMF was added KOtBu (2 eq) at 0° C. Theresulted mixture was warmed to RT and stirred overnight. The reactionmixture was diluted with EtAOc, washed with water, dried over Na₂SO₄,and concentrated in vacuo to give crude product which was used in thenext step without further purification.

Step 4: Preparation of (R) α-substituted cyclohexenecarboxylic acid

A solution of (R) α-methyl ether from Step 3 in 1,4-dioxane, MeOH and 1NNaOH (2:1:1) was stirred at 50° C. The reaction mixture was purified byreverse phase preparative HPLC to give the final product.

Example 20 Preparation of(R)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-2-yloxy)methyl)cyclohex-3-ene-1-carboxylicacid

Step 1-2: General Procedure B Step 3. Preparation of ethyl(R)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-2-yloxy)methyl)cyclohex-3-ene-1-carboxylate

The title compound was prepared as a solid, following the proceduredescribed in General procedure B step 3, using 2-chloropyridine asreactant. LC/MS m/z 830.55 (M+H)⁺, 3.56 min (LCMS Method 5).

Step 4.(R)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-2-yloxy)methyl)cyclohex-3-ene-1-carboxylicacid was prepared in 41% yield (2 steps) as a solid, following theprocedure described in General procedure B step 4 for 6 h, using ethyl(R)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-2-yloxy)methyl)cyclohex-3-ene-1-carboxylateas reactant. ¹H NMR (500 MHz, CHLOROFORM-d) δ 8.21 (dd, J=5.3, 1.4 Hz,1H), 7.70 (ddd, J=8.6, 7.0, 2.0 Hz, 1H), 6.98 (ddd, J=7.1, 5.3, 0.8 Hz,1H), 6.84 (d, J=8.4 Hz, 1H), 5.37 (br. s., 1H), 5.20 (dd, J=6.1, 1.7 Hz,1H), 4.78 (s, 1H), 4.71 (s, 1H), 4.49 (d, J=9.9 Hz, 1H), 4.45 (d, J=9.9Hz, 1H), 3.38-3.31 (m, 1H), 3.25-3.00 (m, 9H), 2.98-2.85 (m, 2H), 2.79(dt, J=10.9, 5.6 Hz, 1H), 2.70-2.62 (m, 1H), 2.28-1.86 (m, 11H),1.76-1.07 (m, 16H), 1.70 (s, 3H), 1.16 (s, 3H), 1.04 (s, 3H), 0.96 (s,3H), 0.93 (s, 3H), 0.87 (s, 3H). LC/MS m/z 802.45 (M+H)⁺, 3.34 min (LCMSMethod 5).

Example 21 Preparation of(R)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyrimidin-4-yloxy)methyl)cyclohex-3-ene-1-carboxylicacid

Step 1-2: General Procedure B Step 3. Preparation of ethyl(R)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyrimidin-4-yloxy)methyl)cyclohex-3-ene-1-carboxylate

The title compound was prepared as a solid, following the proceduredescribed in General procedure B step 3, using 4-chloropyrimidine asreactant. LC/MS m/z 831.55 (M+H)⁺, 3.45 min (LCMS Method 5).

Step 4.(R)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyrimidin-4-yloxy)methyl)cyclohex-3-ene-1-carboxylicacid was prepared in 31% yield (2 steps) as a solid, following theprocedure described in General procedure B step 4 for 4 h, using ethyl(R)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyrimidin-4-yloxy)methyl)cyclohex-3-ene-1-carboxylateas reactant. ¹H NMR (500 MHz, CHLOROFORM-d) δ 8.95 (s, 1H), 8.57 (d,J=6.4 Hz, 1H), 6.96 (dd, J=6.3, 0.9 Hz, 1H), 5.38 (br. s., 1H), 5.21 (d,J=4.6 Hz, 1H), 4.78 (s, 1H), 4.70 (s, 1H), 4.66 (s, 2H), 3.37-3.31 (m,1H), 3.23-3.01 (m, 9H), 2.97-2.86 (m, 2H), 2.80 (dt, J=10.6, 5.6 Hz,1H), 2.69-2.62 (m, 1H), 2.30-1.87 (m, 11H), 1.76-1.01 (m, 16H), 1.70 (s,3H), 1.16 (s, 3H), 1.04 (s, 3H), 0.95 (s, 3H), 0.93 (s, 3H), 0.87 (s,3H). LC/MS m/z 725.50 (M+H)⁺, 3.23 min (LCMS Method 5).

Example 22 Preparation of(R)-1-(((3-chloropyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylicacid

Step 1-2: General Procedure B Step 3. Preparation of ethyl(R)-1-(((3-chloropyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylate

The title compound was prepared as a solid, following the proceduredescribed in General procedure B step 3, using 2,3-dichloropyridine asreactant. LC/MS m/z 864.45 (M+H)⁺, 3.83 min (LCMS Method 5).

Step 4.(R)-1-(((3-chloropyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylicacid was prepared in 69% yield (2 steps) as a solid, following theprocedure described in General procedure B step 4 for 6 h, using ethyl(R)-1-(((3-chloropyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylateas reactant. ¹H NMR (500 MHz, CHLOROFORM-d) δ 8.04 (dd, J=4.9, 1.7 Hz,1H), 7.64 (dd, J=7.6, 1.5 Hz, 1H), 6.87 (dd, J=7.6, 5.0 Hz, 1H), 5.38(br. s., 1H), 5.21 (d, J=4.6 Hz, 1H), 4.77 (s, 1H), 4.71 (s, 1H), 4.54(d, J=10.4 Hz, 1H), 4.51 (d, J=10.2 Hz, 1H), 3.43-3.36 (m, 1H),3.25-3.01 (m, 9H), 2.99-2.87 (m, 2H), 2.75 (td, J=10.9, 5.7 Hz, 1H),2.69-2.62 (m, 1H), 2.30-1.85 (m, 11H), 1.76-1.07 (m, 16H), 1.69 (s, 3H),1.16 (s, 3H), 1.04 (s, 3H), 0.95 (s, 3H), 0.93 (s, 3H), 0.87 (s, 3H).LC/MS m/z 836.45 (M+H)⁺, 3.48 min (LCMS Method 5).

Example 23 Preparation of(R)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylicacid

Step 1-2: General Procedure B Step 3. Preparation of ethyl(R)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 97% yield as a solid, following theprocedure described in General procedure B step 3, using2-fluoronicotinonitrile as reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ8.33 (dd, J=5.0, 2.0 Hz, 1H), 7.87 (dd, J=7.4, 1.9 Hz, 1H), 7.00-6.95(m, 1H), 5.37 (br. s., 1H), 5.19 (d, 7-4.8 Hz, 1H), 4.71 (d, J=2.0 Hz,1H), 4.60 (s, 1H), 4.57-4.53 (m, 2H), 4.18 (qd, J=7.2, 2.6 Hz, 2H),3.12-2.99 (m, 8H), 2.76-2.41 (m, 6H), 2.28-0.90 (m, 27H), 1.69 (s, 3H),1.27 (t, J=7.2 Hz, 3H), 1.06 (s, 3H), 0.97 (s, 3H), 0.96 (s, 3H), 0.93(s, 3H), 0.86 (s, 3H). LC/MS m/z 855.60 (M+H)⁺, 4.03 min (LCMS Method2).

Step 4.(R)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylicacid was prepared in 67% yield as a solid, following the proceduredescribed in General procedure B step 3 at RT for 2 days, using ethyl(R)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylateas reactant. ¹H NMR (400 MHz, METHANOL-d₄) δ 8.37 (dd, J=5.0, 2.0 Hz,1H), 8.06 (dd, J=7.5, 1.8 Hz, 1H), 7.10 (dd, J=7.5, 5.0 Hz, 1H), 5.37(br. s., 1H), 5.22 (dd, J=6.0, 1.5 Hz, 1H), 4.85 (s, 1H), 4.76 (t, J=1.5Hz, 1H), 4.63-4.55 (m, 2H), 3.27-3.07 (m, 11H), 2.91 (ddd, J=14.4, 10.0,4.6 Hz, 1H), 2.79-2.61 (m, 2H), 2.32-1.09 (m, 27H), 1.77 (s, 3H), 1.17(s, 3H), 1.12 (s, 3H), 1.00 (s, 3H), 0.97 (s, 3H), 0.93 (s, 3H). LC/MSm/z 827.60 (M+H)⁺, 3.70 min (LCMS Method 2).

Example 24 Preparation of(R)-1-(((3-carbamoylpyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylicacid

The title compound was a side product formed during Step 4 of thepreparation of ethyl(R)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylate.The material was isolated in 14% yield as a solid. ¹H NMR (400 MHz,METHANOL-d₄) δ 8.40 (dd, J=7.8, 2.0 Hz, 1H), 8.29 (dd, J=5.0, 2.0 Hz,1H), 7.13 (dd, J=7.7, 4.9 Hz, 1H), 5.37 (br. s., 1H), 5.21 (d, J=4.5 Hz,1H), 4.83 (s, 1H), 4.72 (s, 1H), 4.64 (d, J=10.3 Hz, 1H), 4.53 (d,J=10.5 Hz, 1H), 3.28-3.03 (m, 11H), 3.01-2.90 (m, 1H), 2.84-2.68 (m,2H), 2.37-1.06 (m, 27H), 1.75 (s, 3H), 1.18 (s, 3H), 1.10 (s, 3H), 0.99(s, 3H), 0.96 (s, 3H), 0.92 (s, 3H). LC/MS m/z 845.60 (M+H)⁺, 3.66 min(LCMS Method 2).

General Procedure C: Preparation of (S) α-SubstitutedCyclohexenecarboxylic Acid Derivatives.

Step 1. Preparation of((S)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(ethoxycarbonyl)cyclohex-3-en-1-yl)methylbenzoate

The title compound was prepared in 86% of yield as a solid, followingthe procedure described in General procedure B step 1, using(S)-(1-(ethoxycarbonyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-en-1-yl)methylbenzoate instead of(R)-(1-(ethoxycarbonyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-en-1-yl)methylbenzoate as the reactant. LC/MS m/z 857.50 (M+H)⁺, 3.055 min (LCMSMethod 3). ¹H NMR (400 MHz, CHLOROFORM-d) δ 8.07-7.90 (m, 2H), 7.64-7.52(m, 1H), 7.49-7.37 (m, 2H), 5.37 (br. s., 1H), 5.21 (dd, J=6.0, 1.8 Hz,1H), 4.72 (d, J=1.8 Hz, 1H), 4.61 (d, J=1.3 Hz, 1H), 4.52-4.37 (m, 2H),4.25-4.16 (m, 2H), 3.15-3.00 (m, 8H), 2.78-2.53 (m, 5H), 2.51-2.42 (m,1H), 2.34-2.23 (m, 1H), 1.70 (s, 3H), 1.07 (s, 3H), 0.99 (s, 3H), 0.97(s, 3H), 0.93 (s, 3H), 0.87 (s, 3H), 2.22-0.80 (m, 29H).

Step 2. Preparation of ethyl(S)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(hydroxymethyl)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 94% of yield as a solid, followingthe procedure described in General procedure B step 2, using((S)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(ethoxycarbonyl)cyclohex-3-en-1-yl)methylbenzoate instead of ((R)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(ethoxycarbonyl)cyclohex-3-en-1-yl)methylbenzoate as the reactant. LC/MS m/z 753.55 (M+H)⁺, 2.754 min (LCMSMethod 3). ¹H NMR (400 MHz, CHLOROFORM-d) δ 5.30 (s, 1H), 5.16 (d, J=5.0Hz, 1H), 4.72 (s, 1H), 4.61 (s, 1H), 4.23-4.12 (m, 2H), 3.67 (s, 2H),3.28-2.65 (m, 13H), 2.54 (d, J=16.1 Hz, 1H), 1.68 (s, 3H), 1.09 (s, 3H),0.98 (s, 3H), 0.96 (s, 3H), 0.89 (s, 3H), 0.85 (s, 3H), 2.23-0.78 (m,30H).

Step 3. Preparation of ethyl(S)-1-((aryloxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylate

To a solution of ethyl(S)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(hydroxymethyl)cyclohex-3-ene-1-carboxylate(1 eq) in DMF at −78° C. was added KOtBu (2 eq). The resulted mixturewas stirred for 20 minutes before the addition of Ar—X (2 eq). Then thereaction was warmed to RT and stirred overnight. The reaction mixturewas diluted with EtAOc, washed with water, dried over Na₂SO₄, andconcentrated in vacuo to give crude product which was either used innext step without further purification or purified by silica gelchromatography using ethyl ac etate/hexanes as eluents.

Step 4. Preparation of(S)-1-((aryloxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylicacid

A solution of ethyl(S)-1-((aryloxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylatefrom Step 3 in 1,4-dioxane, MeOH and 1N NaOH (2:1:1) was stirred at 50°C. for 2-18 hours. The reaction mixture was then purified by reversephase preparative HPLC to give the final product.

Example 25 Preparation of(S)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylicacid

Step 1-2: General Procedure C Step 1-2 Step 3. Preparation of ethyl(S)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylate

The title compound was prepared as a solid, following the proceduredescribed in General procedure C step 3, using 2-chloronicotinonitrileas the reactant. LC/MS m/z 855.50 (M+H)⁺, 3.004 min (LCMS Method 3).

Step 4.(S)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylicacid was prepared in 29% yield (over 2 steps) as a solid, following theprocedure described in General procedure C step 4 for 7 h, using ethyl(S)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylateas the reactant. LC/MS: m/e 827.50 (M+H)⁺, 3.393 min (LCMS Method 7). ¹HNMR (400 MHz, METHANOL-d₄) δ 8.40 (dd, J=5.1, 1.9 Hz, 1H), 8.08 (dd,J=7.5, 2.0 Hz, 1H), 7.12 (dd, J=7.5, 5.0 Hz, 1H), 5.39 (br. s., 1H),5.25-5.21 (m, 1H), 4.85 (s, 1H), 4.76 (s, 1H), 4.62 (d, J=10.3 Hz, 1H),4.58 (d, J=10.3 Hz, 1H), 3.31-3.18 (m, 8H), 3.16-3.12 (m, 2H), 3.12-3.07(m, 1H), 3.02-2.87 (m, 1H), 2.80 (td, J=11.0, 5.5 Hz, 1H), 2.73-2.63 (m,2H), 2.37-2.27 (m, 1H), 2.26-2.01 (m, 8H), 1.97-1.91 (m, 1H), 1.88-1.75(m, 2H), 1.78 (s, 3H), 1.72-1.44 (m, 10H), 1.42-1.31 (m, 1H), 1.20 (s,3H), 1.27-1.09 (m, 3H), 1.13 (s, 3H), 1.02 (s, 3H), 0.98 (s, 3H), 0.95(s, 3H).

Alternatively, Example 28 can be prepared using the following procedure:

Step 1: Preparation of(S)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(hydroxymethyl)cyclohex-3-enecarboxylicacid, HCl. To a flask containing a suspension of(R)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(fluoromethyl)cyclohex-3-enecarboxylicacid (4.08 g, 5.61 mmol) prepared as described in WO 2015157483 in1,4-dioxane (50.0 mL) was added tetrabutylaminonium hydroxide (55% inwater) (26.5 g, 56.1 mmol). The flask was attached to reflux condensorand was heated in an oil bath at 100° C. After 8.5 days of heating,LC/MS showed the reaction was complete. The mixture was cooled to rt andwas transferred to a graduated addition funnel. Upon standing in theaddition funnel, two distinct layers formed. The bottom layer containingthe product was split in half based on the graduation of the funnel.Half of the material was made acidic by adding 1N HCl. The solids thatformed were collected by filtration and were washed with water. Thesolids were then triturated with ether and collected by filtration. Thesolids were washed with ether then allowed to dry on the filter paper.The title product was isolated as a white solid (1-95 g, 2.56 mmol,45.6% yield, 91% if calculated as half of the mixture). LCMS: m/e 725.4(M+H)⁺, 1.15 min (method 16).

Step 2. To a suspension of(S)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(hydroxymethyl)cyclohex-3-enecarboxylicacid, HCl (1.95 g, 2.56 mmol) in THF (30 mL) was added KHMDS (0.91M inTHF) (9.0 mL, 8.19 mmol). The mixture was stirred for 5 minutes, then2-fluoronicotinonitrile (1.0 g, 8.19 mmol) was added. After 2.5 h analiquot was removed. LC/MS showed the reaction was complete. Thereaction mixture was diluted with 1N HCl (30 mL) then was extracted withethyl acetate (3×75 mL). The organic layers were washed with sat. aq.NaCl, and dried over magnesium sulfate. The drying agent was removed byfiltration. The drying agent did not filter well, so it is likely thatsolid precipitated while standing at rt, so the solid filter cake wasstirred with ethyl acetate, then with dichloromethane, then filteredagain. The combined filtrates were concentrated under reduced pressure.The residue was triturated with ether and the solids that formed werecollected by filtration and washed with ether. The residue was dissolvedin methanol and was purified by reverse phase chromatography using a 275g Isco Redisep gold C18 column and a 20% B-80% A to 100% B gradientwhere A was 90% water, 10% acetonitrile with 0.1% TFA buffer and B was10% water, 90% acetonitrile with 0.1% TFA buffer. The fractionscontaining the product were combined and concentrated under reducedpressure to give(S)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid, TFA (1.50 g, 1.59 mmol, 62%) as a white solid. LCMS: m/e 827.4(M+H)⁺, 1.32 min (method 16).

Example 26 Preparation of(S)-1-(((3-carbamoylpyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylicacid

The title compound was prepared as a side product in 7% yield (over 2steps) as a solid, following the procedure described in Generalprocedure C Step 4 for 7 h, using ethyl(S)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylateas the reactant. LC/MS: m/e 845.55 (M+H—H₂O)⁺, 3.349 min (LCMS Method7). ¹H NMR (400 MHz, METHANOL-d₄) δ 8.42 (dd, J=7.5, 2.0 Hz, 1H), 8.31(dd, J=4.8, 2.0 Hz, 1H), 7.16 (dd, J=7.5, 5.0 Hz, 1H), 5.40 (br. s.,1H), 5.24 (d, J=4.5 Hz, 1H), 4.86 (br. s., 1H), 4.76 (s, 1H), 4.66-4.62(d, J=10.5 Hz, 1H), 4.57-4.53 (d, J=10.5 Hz, 1H), 3.30-3.17 (m, 7H),3.12 (d, J=17.3 Hz, 3H), 2.96-2.92 (m, 1H), 2.81-2.71 (m, 2H), 2.45-2.30(m, 1H), 2.25-2.12 (m, 5H), 2.12-2.00 (m, 3H), 1.92-1.67 (m, 6H), 1.78(s, 3H), 1.67-1.41 (m, 10H), 1.26-1.06 (m, 3H), 1.20 (s, 3H), 1.13 (s,3H), 1.02 (s, 3H), 0.99 (s, 3H), 0.95 (s, 3H).

Example 27 Preparation of(R)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((3-(methoxycarbonyl)pyridin-2-yl)oxy)methyl)cyclohex-3-ene-1-carboxylicacid

The title compound was prepared as a side product in 0.6% yield (over 2steps) as a solid, following the procedure described in Generalprocedure B step 4 for 15 h, using ethyl(R)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylateas the reactant. LC/MS: m/e 860.65 (M+H)⁺, 2.93 min (LCMS Method 7). ¹HNMR (400 MHz, METHANOL-d₄) δ 8.30 (dd, J=5.0, 2.0 Hz, 1H), 8.17 (dd,J=7.5, 2.0 Hz, 1H), 7.06 (dd, J=7.5, 5.0 Hz, 1H), 5.39 (br. s., 1H),5.25-5.21 (m, 1H), 4.85 (s, 1H), 4.75 (s, 1H), 4.57-4.47 (m, 2H), 3.89(s, 3H), 3.30-3.17 (m, 8H), 3.16-3.07 (m, 3H), 3.02-2.90 (m, 1H), 2.81(td, J=11.0, 5.4 Hz, 1H), 2.73-2.63 (m, 1H), 2.36-2.00 (m, 9H),2.00-1.90 (m, 1H), 1.90-1.75 (m, 3H), 1.77 (s, 3H), 1.75-1.34 (m, 12H),1.26-1.09 (m, 2H), 1.20 (s, 3H), 1.12 (s, 3H), 1.01 (s, 3H), 0.98 (s,3H), 0.95 (s, 3H).

Example 28 Preparation of(S)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((6-methoxypyridin-2-yl)oxy)methyl)cyclohex-3-ene-1-carboxylicacid

Step 1-2: General Procedure C Step 1-2 Step 3. Preparation of ethyl(S)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((6-fluoropyridin-2-yl)oxy)methyl)cyclohex-3-ene-1-carboxylate

The title compound was prepared as a solid, following the proceduredescribed in General procedure C step 3, using 2,6-difluoropyridine asthe reactant. LC/MS m/z 848.50 (M+H)⁺, 3.031 min (LCMS Method 3).

Step 4.(S)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((6-methoxypyridin-2-yl)oxy)methyl)cyclohex-3-ene-1-carboxylicacid was prepared in 3.7% yield (over 2 steps) as a solid, following theprocedure described in General procedure C step 4 for 15 h, using ethyl(S)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((6-fluoropyridin-2-yl)oxy)methyl)cyclohex-3-ene-1-carboxylateas the reactant. LC/MS: m/e 832.50 (M+H)⁺, 3.267 min (LCMS Method 7).NMR (500 MHZ, METHANOL-d₄) δ 7.54 (t, J=7.9 Hz, 1H), 6.32 (d, J=2.9 Hz,1H), 6.30 (d, J=2.9 Hz, 1H), 5.38 (br. s., 1H), 5.29-5.15 (m, 1H), 4.85(s, 1H), 4.76 (s, 1H), 4.485-4.345 (m, 2H), 3.89 (s, 3H), 3.30-3.17 (m,8H), 3.17-3.07 (m, 3H), 2.94 (ddd, J=14.5, 10.2, 4.7 Hz, 1H), 2.78 (td,J=1.0, 5.4 Hz, 1H), 2.67-2.60 (m, 1H), 2.30 (d, J=18.2 Hz, 1H),2.22-2.09 (m, 3H), 2.09-2.00 (m, 2H), 1.96-1.66 (m, 8H), 1.78 (s, 3H),1.66-1.43 (m, 10H), 1.43-1.29 (m, 2H), 1.29-1.09 (m, 1H), 1.19 (s, 3H),1.13 (s, 3H), 1.01 (s, 3H), 0.98 (s, 3H), 0.95 (s, 3H).

Example 29 Preparation of(S)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((6-fluoropyridin-2-yl)oxy)methyl)cyclohex-3-ene-1-carboxylicacid

The title compound was prepared in 69.8% of yield (2 steps) as a solid,following the procedure described in General procedure C step 4 for 7 h,using ethyl(S)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((6-fluoropyridin-2-yl)oxy)methyl)cyclohex-3-ene-1-carboxylateas the reactant. LC/MS: m/e 820.45 (M+H)⁺, 3.136 min (LCMS Method 7).NMR (500 MHz, ACETONE-d₆) δ 7.85 (q, J=8.1 Hz, 1H), 6.71 (dd, J=8.0, 1.3Hz, 1H), 6.61 (dd, J=7.8, 2.3 Hz, 1H), 5.42-5.35 (m, 1H), 5.23 (dd,J=6.2, 1.8 Hz, 1H), 4.79 (d, J=1.2 Hz, 1H), 4.68 (d, J=1.4 Hz, 1H), 4.46(d, J=10.2 Hz, 1H), 4.41 (d, J=10.2 Hz, 1H), 3.43-3.24 (m, 8H),3.23-3.12 (m, 5H), 3.12-3.05 (m, 3H), 3.02 (td, J=10.8, 5.7 Hz, 1H),2.70-2.61 (m, 1H), 2.38-2.16 (m, 4H), 2.17-2.01 (m, 3H), 1.95-1.84 (m,2H), 1.84-1.68 (m, 2H), 1.74 (s, 3H), 1.64 (d, J=16.8 Hz, 1H), 1.61-1.42(m, 8H), 1.40-1.22 (m, 1H), 1.26 (s, 3H), 1.23-1.11 (m, 2H), 1.13 (s,3H), 1.02 (s, 3H), 0.97 (s, 3H), 0.94 (s, 3H).

Example 30 Preparation of(S)-1-(((4-carbamoylpyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylicacid

Step 1-2: General Procedure C Step 1-2 Step 3. Preparation of ethyl(S)-1-(((4-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylate

The title compound was prepared as a solid, following the proceduredescribed in General procedure C step 3, using2-fluoroisonicotinonitrile as the reactant. LC/MS m/z 855.50 (M+H)⁺,3.048 min (LCMS Method 3).

Step 4.(S)-1-(((4-carbamoylpyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylicacid was prepared in 30.5% yield (over 2 steps) as a solid, followingthe procedure described in General procedure C step 4 for 7 h, usingethyl(S)-1-(((4-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylateas the reactant. LC/MS: m/e 845.55 (M+H)⁺, 3.048 min (LCMS Method 7). ¹HNMR (400 MHz, CHLOROFORM-d) δ 8.02 (d, J=5.3 Hz, 1H), 7.12 (d, J=5.3 Hz,1H), 6.97 (s, 1H), 5.16 (br. s., 1H), 5.00 (d, J=5.5 Hz, 1H), 4.58 (br.s., 1H), 4.48 (br. s., 1H), 4.35-4.18 (m, 2H), 3.25-2.65 (m, 18H), 2.47(d, J=17.1 Hz, 1H), 2.14-1.64 (m, 10H), 1.52-1.48 (m, 2H), 1.50 (s, 3H),1.45-1.03 (m, 10H), 0.98 (s, 3H), 0.88-0.84 (m, 2H), 0.86 (s, 3H), 0.78(s, 3H), 0.73 (s, 3H), 0.68 (s, 3H).

Example 31 Preparation of(S)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-2-yloxy)methyl)cyclohex-3-ene-1-carboxylicacid

Step 1-2: General Procedure C Step 1-2 Step 3. Preparation of ethyl(S)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-2-yloxy)methyl)cyclohex-3-ene-1-carboxylate

The title compound was prepared as a solid, following the proceduredescribed in General procedure C step 3, using 2-bromopyridine as thereactant. LC/MS m/z M+1=830.55. 2.822 min (LCMS Method 3).

Step 4.(S)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-2-yloxy)methyl)cyclohex-3-ene-1-carboxylicacid was prepared in 22.9% yield (over 2 steps) as a solid, followingthe procedure described in General procedure C step 4 for 7 h, usingethyl(S)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-2-yloxy)methyl)cyclohex-3-ene-1-carboxylateas the reactant. LC/MS: m/e 802.45 (M+H)⁺, 2.824 min (LCMS Method 3). ¹HNMR (400 MHz, CHLOROFORM-d) δ 8.21 (dd, J=5.1, 1.4 Hz, 1H), 7.69 (ddd,J=8.6, 7.0, 1.8 Hz, 1H), 6.97 (td, J=6.2, 0.9 Hz, 1H), 6.83 (d, J=8.5Hz, 1H), 5.38 (br. s., 1H), 5.21 (d, J=4.5 Hz, 1H), 4.80 (s, 1H), 4.72(s, 1H), 4.51 (d, J=10.0 Hz 1H), 4.46 (d, J=10.0 Hz 1H), 3.37-3.34 (m,1H), 3.25-3.10 (m, 7H), 3.10-3.01 (m, 2H), 3.00-2.87 (m, 2H), 2.82 (dt,J=10.9, 5.6 Hz, 1H), 2.73 (d, J=15.3 Hz, 1H), 2.35-2.13 (m, 4H),2.13-1.88 (m, 7H), 1.81-1.67 (m, 2H), 1.71 (s, 3H), 1.66-1.26 (m, 13H),1.18 (s, 3H), 1.13-1.03 (m, 1H), 1.06 (s, 3H), 0.99 (s, 3H), 0.94 (s,3H), 0.89 (s, 3H).

Example 32 Preparation of(S)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyrazin-2-yloxy)methyl)cyclohex-3-ene-1-carboxylicacid

Step 1-2: General Procedure C Step 1-2 Step 3. Preparation of ethyl(S)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyrazin-2-yloxy)methyl)cyclohex-3-ene-1-carboxylate

The title compound was prepared as a solid, following the proceduredescribed in General procedure C step 3, using 2-fluoropyrazine as thereactant. LC/MS m/z M+1=831.55. 2.922 min (LCMS Method 3).

Step 4.(S)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyrazin-2-yloxy)methyl)cyclohex-3-ene-1-carboxylicacid was prepared in 77.0% yield (over 2 steps) as a solid, followingthe procedure described in General procedure C step 4 for 9 h, usingethyl(S)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyrazin-2-yloxy)methyl)cyclohex-3-ene-1-carboxylateas the reactant. LC/MS: m/e 803.42 (M+H)⁺, 2.38 min (LCMS Method 1). ¹HNMR (400 MHz, CHLOROFORM-d) δ 8.27 (br. s., 2H), 8.17 (br. s., 1H), 5.39(br. s., 1H), 5.22 (d, J=4.8 Hz, 1H), 4.79 (s, 1H), 4.73 (s, 1H),4.62-4.48 (dd, J=10.5, 17.3 Hz, 2H), 3.44-3.32 (m, 1H), 3.30-2.89 (m,11H), 2.84-2.64 (m, 2H), 2.38-1.83 (m, 11H), 1.83-1.67 (m, 2H), 1.71 (s,3H), 1.68-1.37 (m, 10H), 1.38-1.22 (m, 2H), 1.16 (s, 3H), 1.13-1.03 (m,2H), 1.06 (s, 3H), 0.98 (s, 3H), 0.93 (s, 3H), 0.89 (s, 3H).

General Procedure D: Preparation of α-Pyridin-2-YloxyCyclohexenecarboxylic Acid Derivatives.

Step 1. Preparation of ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-2-yloxy)methyl)cyclohex-3-ene-1-carboxylate

A mixture of (1R,3aS,5aR,5bR,7aR,11aR,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yltrifluoromethanesulfonate (1 eq), ethyl1-((pyridin-2-yloxy)methyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-ene-1-carboxylate(1 eq), Na₂CO₃ (3 eq) and Pd(Ph₃P)₄ (0.06 eq) in 1,4-dioxane and H₂O(4:1), was flushed with nitrogen, sealed and heated at 70° C. for 2 h.The reaction mixture was diluted with EtOAc, washed with brine, driedover Na₂SO₄, and concentrated in vacuo. The crude product was purifiedby silica gel column eluted with 0-55% ethyl acetate/hexanes to give thedesired product (57% yield) as a solid. ¹H NMR (400 MHz, CHLOROFORM-d) δ8.13 (dd, J=5.0, 1.5 Hz, 1H), 7.58-7.52 (m, 1H), 6.86 (ddd, J=7.2, 5.1,0.8 Hz, 1H), 6.72 (d, J=8.5 Hz, 1H), 5.35 (br. s., 1H), 5.19 (d, J=5.8Hz, 1H), 4.73 (d, J=2.3 Hz, 1H), 4.60 (dd, J=2.3, 1.3 Hz, 1H), 4.48-4.37(m, 2H), 4.18-4.11 (m, 2H), 2.70-2.62 (m, 1H), 2.54 (td, J=10.9, 5.3 Hz,1H), 2.29-0.84 (m, 27H), 1.69 (s, 3H), 1.20 (t, J=7.2 Hz, 3H), 1.07 (s,3H), 0.96 (s, 3H), 0.97-0.91 (m, 6H), 0.86 (s, 3H). LC/MS m/z 669.60(M+H)⁺, 2.82 min (LCMS Method 3).

Step 2: Preparation of C-17 Amine Derivative

To a solution of ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-2-yloxy)methyl)cyclohex-3-ene-1-carboxylate(1 eq) and aldehyde (2 eq) in DCE was added titanium (IV) isopropoxide(2 eq). The mixture was stirred at RT for 1 h. Sodiumtriacetoxyborohydride (2 eq) was added and the mixture was stirred at RTovernight. The reaction was quenched with saturated aqueous Na₂CO₃. Theresulting slurry was extracted with dichloromethane, washed with brine,dried over Na₂SO₄, and concentrated in vacuo. The crude product waspurified by silica gel column eluted with ethyl acetate/hexanes to givethe desired product.

Step 3: Preparation of Carboxylic Acid

A solution of the ester from step 2 in 1,4-dioxane, MeOH and 1N NaOH(2:1:1) was stirred at 60-70° C. The reaction mixture was purified byreverse phase preparative HPLC to give the final product.

Example 33 Preparation of4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(4-hydroxy-1,1-dioxidotetrahydro-2H-thiopyran-4-yl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-2-yloxy)methyl)cyclohex-3-ene-1-carboxylicacid

Step 1: General Procedure D Step 1 Step 2. Preparation of ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(4-hydroxy-1,1-dioxidotetrahydro-2H-thiopyran-4-yl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-2-yloxy)methyl)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 78% yield as a solid, following theprocedure described in general procedure D step 2, using2-(4-hydroxy-1,1-dioxidotetrahydro-2H-thiopyran-4-yl)acetaldehyde asreactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 8.12 (dd, J=5.0, 1.5 Hz, 1H),7.57-7.52 (m, 1H), 6.85 (ddd, J=7.0, 5.1, 0.9 Hz, 1H), 6.72 (d, J=8.5Hz, 1H), 5.34 (br. s., 1H), 5.18 (d, J=5.8 Hz, 1H), 4.73 (d, J=1.8 Hz,1H), 4.61 (s, 1H), 4.48-4.36 (m, 2H), 4.18-4.08 (m, 4H), 3.57-3.43 (m,2H), 2.91-2.61 (m, 5H), 2.50 (td, J=10.7, 5.5 Hz, 1H), 2.24-0.88 (31H),1.68 (s, 3H), 1.19 (t, J=7.2 Hz, 3H), 1.03 (s, 3H), 0.96 (s, 3H),0.95-0.90 (m, 6H), 0.85 (s, 3H). LC/MS m/z 845.60 (M+H)⁺, 3.59 min (LCMSMethod 4).

Step 3.4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(4-hydroxy-1,1-dioxidotetrahydro-2H-thiopyran-4-yl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-2-yloxy)methyl)cyclohex-3-ene-1-carboxylicacid was prepared in 76% yield as a solid, following the proceduredescribed in general procedure D step 3 at 60° C. for 12 h, using ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(4-hydroxy-1,1-dioxidotetrahydro-2H-thiopyran-4-yl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-2-yloxy)methyl)cyclohex-3-ene-1-carboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 8.29 (br. s., 1H), 7.96(t, J=7.0 Hz, 1H), 7.17 (br. s., 1H), 7.04 (d, J=8.3 Hz, 1H), 5.35 (br.s., 1H), 5.18 (d, J=5.3 Hz, 1H), 4.75 (s, 1H), 4.69 (s, 1H), 4.53-4.39(m, 2H), 3.58-3.35 (m, 2H), 3.21 (br. s., 2H), 2.97-2.84 (m, 2H),2.76-2.62 (m, 2H), 2.58-2.44 (m, 1H), 2.34-1.04 (m, 32H), 1.68 (s, 3H),1.07 (s, 3H), 1.02 (s, 3H), 0.95-0.91 (m, 6H), 0.87 (s, 3H). LC/MS m/z817.55 (M+H)⁺, 5.51 min (LCMS Method 4).

General Procedure E. Preparation of α-Substituted CyclohexenecarboxylicAcid Derivatives Via Alkylation of α-Methyl Alcohol.

Step 1. Preparation of(4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(ethoxycarbonyl)cyclohex-3-en-1-yl)methylbenzoate

A mixture of(1R,3aS,5aR,5bR,7aR,11aR,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yltrifluoromethanesulfonate (1 eq), (1-(ethoxycarbonyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-en-1-yl)methylbenzoate (1.05 eq), Na₂CO₃H₂O (3 eq) and Pd(Ph3P)₄ (0.06 eq) in1,4-dioxane and H₂O (4:1) was flushed with nitrogen, sealed and heatedat 70° C. for 2 h. The reaction mixture was concentrated in vacuo, andthe residue was partitioned between EtOAc and H₂O. The separated aqueouslayer was extracted with EtOAc. The combined organic layers were washedwith brine dried over Na₂SO₄, and concentrated in vacuo. The crudeproduct was purified by silica gel column eluted with 0-60% ethylacetate/hexanes to give the desired product as a solid (67% yield). ¹HNMR (400 MHz, CHLOROFORM-d) δ 8.01 (dd, J=8.2, 1.1 Hz, 2H), 7.59-7.53(m, 1H), 7.46-7.40 (m, 2H), 5.36 (br. s., 1H), 5.20 (d, J=5.5 Hz, 1H),4.71 (d, J=2.0 Hz, 1H), 4.59 (s, 1H), 4.48-4.39 (m, 2H), 4.21-4.14 (m,2H), 3.12-2.98 (m, 8H), 2.73-2.53 (m, 5H), 2.50-2.42 (m, 1H), 2.31-0.81(m, 27H), 1.69 (s, 3H), 1.22 (t, J=7.2 Hz, 3H), 1.06 (s, 3H), 0.96 (s,3H), 0.98-0.92 (m, 6H), 0.86 (s, 3H). LC/MS: m/e 857.50 (M+H)⁺, 2.91 min(LCMS Method 3).

Step 2:. Preparation of ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(hydroxymethyl)cyclohex-3-ene-1-carboxylate

A suspension of4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(ethoxycarbonyl)cyclohex-3-en-1-yl)methylbenzoate (1 eq) and 1N NaOH (1 eq) in MeOH and THF was stirred at RT for1 day. The mixture was neutralized with saturated aqueous citric acidand the solvent was removed in vacuo. The residue was taken into EtOAc,washed with brine, dried over Na₂SO₄, and concentrated in vacuo to givethe desired product (99% yield) as a solid without further purification.LC/MS m/z 753.70 (M+H)⁺, 2.85 min (LCMS Method 3).

Step 3. Preparation of α-Substituted Cyclohexenecarboxylic Ester

To a solution ofethyl-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(hydroxymethyl)cyclohex-3-ene-1-carboxylate(1 eq) and Ar—X (2 eq) in DMF was added KOtBu (2 eq). The resultingmixture was warmed to RT and stirred overnight. The reaction mixture wasdiluted with EtAOc, washed with water, dried over Na₂SO₄, andconcentrated in vacuo to give crude product which was used in next stepwithout further purification.

Step 4. Preparation of α-Substituted Cyclohexenecarboxylic Acid

A solution of α-methyl ether from step 4 in 1,4-dioxane, MeOH and 1NNaOH (2:1:1) was stirred at 50° C. The reaction mixture was purified byreverse phase preparative HPLC to give the final product.

Example 34 Preparation of4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyrimidin-2-yloxy)methyl)cyclohex-3-ene-1-carboxylicacid

Step 1-2: General Procedure E Step 3. Preparation of ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyrimidin-2-yloxy)methyl)cyclohex-3-ene-1-carboxylate

The title compound was prepared as crude product, following theprocedure described in general procedure E step 3, using2-bromopyrimidine as reactant. LC/MS m/z 831.60 (M+H)⁺, 2.76 min (LCMSMethod 3).

Step 4.4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyrimidin-2-yloxy)methyl)cyclohex-3-ene-1-carboxylicacid was prepared in 11% yield as a solid, following the proceduredescribed in general procedure E step 4, using ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyrimidin-2-yloxy)methyl)cyclohex-3-enecarboxylateas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 8.62 (d, J=4.8 Hz, 2H),7.10 (t, J=4.9 Hz, 1H), 5.39 (br. s., 1H), 5.21 (d, J=4.5 Hz, 1H), 4.78(s, 1H), 4.72 (s, 1H), 4.64-4.54 (m, 2H), 3.39 (br. d, J=13.1 Hz, 1H),3.27-3.03 (m, 9H), 3.03-2.89 (m, 2H), 2.80-2.70 (m, 1H), 2.33-2.06 (m,4H), 2.06-2.02 (m, 6H), 2.02-1.85 (m, 4H), 1.81-1.67 (m, 2H), 1.71 (s,3H), 1.67-1.37 (m, 10H), 1.37-1.25 (m, 2H), 1.16 (s, 3H), 1.12-1.03 (m,1H), 1.06 (s, 3H), 0.98-0.97 (m, 3H), 0.95-0.94 (m, 3H), 0.89 (s, 3H).LC/MS: m/e 803.50 (M+H)⁺, 2.80 min (LCMS Method 3).

Example 35 Preparation of4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((7-methoxyisoquinolin-1-yl)oxy)methyl)cyclohex-3-ene-1-carboxylicacid

Step 1-2: General Procedure E Step 3. Preparation of ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((7-methoxyisoquinolin-1-yl)oxy)methyl)cyclohex-3-enecarboxylate

The title compound was prepared as crude product, following theprocedure described in general procedure E step 3, using1-chloro-7-methoxyisoquinoline as reactant. LC/MS: m/e 910.65 (M+H)⁺,2.98 min (LCMS Method 3).

Step 4:4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((7-methoxyisoquinolin-1-yl)oxy)methyl)cyclohex-3-ene-1-carboxylicacid was prepared in 39% yield as a solid, following the proceduredescribed in general procedure E step 4, using ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((7-methoxyisoquinolin-1-yl)oxy)methyl)cyclohex-3-enecarboxylateas reactant. LC/MS: m/e 882.60 (M+H)⁺, 2.83 min (LCMS Method 3).

Example 36 Preparation of1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylicacid

Step 1-2: General Procedure E Step 3. Preparation of ethyl1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylate

The title compound was prepared in 41% yield, following the proceduredescribed in general procedure E step 3, using 2-chloronicotinonitrileas reactant. ¹H NMR (400 MHz, CHLOROFORM-d) δ 8.31 (dd, J=5.0, 1.8 Hz,1H), 7.86 (dd, J=7.5, 2.0 Hz, 1H), 6.97 (dd, J=7.4, 5.1 Hz, 1H), 5.34(br. s., 1H), 5.17 (d, J=5.0 Hz, 1H), 4.69 (d, J=1.8 Hz, 1H), 4.57 (br.s., 1H), 4.53 (s, 2H), 4.21-4.12 (m, 2H), 3.10-2.97 (m, 8H), 2.74-2.40(m, 6H), 2.28-0.82 (m, 27H), 1.67 (s, 3H), 1.25 (t, J=7.2 Hz, 3H), 1.04(s, 3H), 0.94 (s, 3H), 0.93-0.88 (m, 6H), 0.84 (s, 3H). LC/MS: m/e855.60 (M+H)⁺, 3.08 min (LCMS Method 7).

Step 4.1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylicacid was prepared in 33% yield, following the procedure described ingeneral procedure E step 3 at RT, using ethyl1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylateas reactant. ¹H NMR (400 MHz, METHANOL-d₄) □ 8.41-8.38 (m, 1H), 8.08(dd, J=7.5, 1.8 Hz, 1H), 7.12 (dd, J=7.7, 5.1 Hz, 1H), 5.39 (br. s.,1H), 5.23 (d, J=4.8 Hz, 1H), 4.85 (s, 1H), 4.75 (s, 1H), 4.63 (dd,J=3.8, 10.5 Hz, 1H), 4.58 (d, J=10.3 Hz, 1H), 3.30-3.17 (m, 8H),3.17-3.07 (m, 3H), 2.99-2.89 (m, 1H), 2.80 (td, J=11.0, 5.4 Hz, 1H),2.72-2.64 (m, 1H), 2.40-2.23 (m, 1H), 2.23-2.15 (m, 2H), 2.15-2.01 (m,7H), 1.99-1.90 (m, 1H), 1.90-1.76 (m, 3H), 1.78 (s, 3H), 1.76-1.64 (m,2H), 1.63-1.41 (m, 9H), 1.41-1.29 (m, 1H), 1.24-1.18 (m, 1H), 1.20 (s,3H), 1.18-1.10 (m, 1H), 1.13 (s, 3H), 1.025-1.015 (m, 3H), 0.98 (s, 3H),0.95 (s, 3H). LC/MS: m/e 827.65 (M+H)⁺, 3.12 min (LCMS Method 7).

General Procedure F. Preparation of α-Substituted CyclohexenecarboxylicAcid Derivatives Via Silyl Carboxylate.

Step 1. Preparation of (1-((benzyloxy)carbonyl)-4-oxocyclohexyl)methylbenzoate

To a solution of benzyl 1-(hydroxymethyl)-4-oxocyclohexanecarboxylate(4.3 g, 16.4 mmol) in pyridine (20 mL) was added benzoic anhydride (4.45g, 19.7 mmol) followed by DMAP (2.00 g, 16.4 mmol). The resultingsolution was stirred at 55° C. for 2 hours. The reaction mixture wasdiluted with 50 mL of ethyl acetate and was washed with 0.5 N HCl topH=4. The organic layer was dried over Na₂SO₄ and concentrated in vacuo.The crude product was purified by silica gel column eluted with 0-50%ethyl acetate/hexanes to give the desired product as an oil (3.3 g,49%).

¹H NMR (400 MHz, CHLOROFORM-d) δ 7.92 (d, J=7.8 Hz, 2H), 7.65-7.54 (m,1H), 7.44-7.37 (m, 2H), 7.35-7.27 (m, 5H), 5.25 (s, 2H), 4.46 (s, 2H),2.63-2.35 (m, 6H), 1.86 (td, J=12.4, 5.0 Hz, 2H).

Step 2. Preparation of(1-((benzyloxy)carbonyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-en-1-yl)methylbenzoate

To a solution of (1-((benzyloxy)carbonyl)-4-oxocyclohexyl)methylbenzoate (4.2 g, 11.5 mmol) and1,1,1-trifluoro-N-phenyl-N-((trifluoromethyl)sulfonyl)-methanesulfonamide(4.5 g, 12.6 mmol) in THF (50 mL) at −78° C. was added KHMDS (1 M inTHF) (14.9 mL, 14.9 mmol). The resulting yellow solution was stirred at−78° C. for 2 h. The reaction was quenched with saturated aqueous NH₄Cl.The mixture was extracted with EtOAc, washed with brine, dried overNa₂SO₄, and concentrated in vacuo. The crude product was purified bysilica gel column eluted with 0-15% ethyl acetate/hexanes to give thedesired triflate as an oil (3.6 g, 63%). ¹H NMR (400 MHz, CHLOROFORM-d)δ 7.92 (d, J=7.8 Hz, 2H), 7.62-7.55 (m, 1H), 7.42 (t, J=7.5 Hz, 2H),7.35-7.27 (m, 5H), 5.80 (br. s., 1H), 5.26-5.14 (m, 2H), 4.50-4.41 (m,2H), 2.90 (dd, J=17.9, 2.4 Hz, 1H), 2.57-2.28 (m, 4H), 2.02-1.91 (m,1H).

Step 3. Preparation of(1-((benzyloxy)carbonyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-en-1-yl)methylbenzoate

A mixture of(1-((benzyloxy)carbonyl)-4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-en-1-yl)methylbenzoate (3.32 g, 6.66 mmol), bis(pinacolato)diboron (1.71 g, 6.73mmol), KOAc (1.64 g, 16.7 mmol)) and PdCl₂(dppf)-CH₂Cl₂ adduct (0.16 g,0.2 mmol) in 1,4-dioxane (30 mL) was flushed with nitrogen, sealed andheated at 70° C. for 20 h. The mixture was diluted with water (150 mL)and extracted with EtOAc (3×125 mL). The combined organic layers werewashed with brine, dried over Na₂SO₄, and concentrated in vacuo. Thecrude product was purified by silica gel column eluted with 0-20% ethylacetate/hexanes to give the desired boronate as an oil (2.2 g, 69%). ¹HNMR (400 MHz, CHLOROFORM-d) δ 7.90 (d, J=8.1 Hz, 2H), 7.58-7.51 (m, 1H),7.42-7.36 (m, 2H), 7.32-7.22 (m, 5H), 6.54 (br. s., 1H), 5.16 (s, 2H),4.48-4.36 (m, 2H), 2.75 (d, J=17.6 Hz, 1H), 2.32-2.19 (m, 3H), 2.07-2.00(m, 1H), 1.92-1.86 (m, 1H), 1.27 (s, 12H). LC/MS: m/e 499.20 (M+Na)⁺,3.10 min (LCMS Method 7).

Step 4. Preparation of(4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-1-(buta-2,3-dien-2-yl)-5a,5b,8,8,11a-pentamethyl-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((benzyloxy)carbonyl)cyclohex-3-en-1-yl)methylbenzoate

A mixture of(1R,3aS,5aR,5bR,7aR,11aR,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yltrifluoromethanesulfonate (2.4 g, 4.3 mmol),(1-((benzyloxy)carbonyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-en-1-yl)methylbenzoate (2.05 g, 4.3 mmol), Na₂CO₃H₂O (1.60 g, 12.9 mmol) and Pd(Ph₃P)₄(0.3 g, 0.26 mmol) in 1,4-dioxane (100 mL) and H₂O (25 mL) was flushedwith nitrogen, sealed and heated at 70° C. for 2 h. The reaction mixturewas diluted with EtOAc, washed with brine, dried over Na₂SO₄, andconcentrated in vacuo. The crude product was purified by silica gelcolumn eluted with 0-55% ethyl acetate/hexanes to give the desired C-3α-substituted cyclohexenecarboxylic ester (1.8 g, 55%). ¹H NMR (400 MHz,CHLOROFORM-d) δ 7.91 (d, J=7.0 Hz, 2H), 7.58-7.51 (m, 1H), 7.42-7.35 (m,2H), 7.33-7.28 (m, 2H), 7.26-7.22 (m, 3H), 5.34 (br. s., 1H), 5.21-5.11(m, 3H), 4.73 (s, 1H), 4.60 (br. s., 1H), 4.51-4.39 (m, 2H), 2.71 (d,J=17.3 Hz, 1H), 2.54 (td, J=10.9, 5.1 Hz, 1H), 2.25-0.92 (m, 27H), 1.69(s, 3H), 1.13-0.85 (m, 15H). LC/MS: m/e 758.70 (M+H)⁺, 3.24 min (LCMSMethod 7).

Step 5. Preparation of(1-((benzyloxy)carbonyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-en-1-yl)methylbenzoate

A suspension of(4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((benzyloxy)carbonyl)cyclohex-3-en-1-yl)methylbenzoate (1.6 g, 2.11 mmol), 4-(2-chloroethyl)thiomorpholine 1,1-dioxidehydrochloride (1.5 g, 6.33 mmol), sodium iodide (0.35 g, 2.32 mmol) andK₃PO₄ (2.24 g, 10.55 mmol) in acetonitrile (20 mL) was flushed with N₂,sealed and heated at 100° C. for 15 h. The reaction mixture was dilutedwith EtOAc (100 mL), washed with water (100 mL), dried over Na₂SO₄, andconcentrated in vacuo. The crude product was purified on silica gelcolumn eluted with 25-60% EtOAc/hexane to give the desired product (1.3g, 67% yield). ¹H NMR (400 MHz, CHLOROFORM-d) δ 7.92 (d, J=7.8 Hz, 2H),7.58-7.51 (m, 1H), 7.43-7.36 (m, 2H), 7.31 (d, J=4.6 Hz, 2H), 7.25 (d,J=4.4 Hz, 3H), 5.35 (br. s., 1H), 5.22-5.12 (m, 3H), 4.71 (s, 1H), 4.60(br. s., 1H), 4.45 (q, J=10.7 Hz, 2H), 3.15-2.99 (m, 8H), 2.78-2.42 (m,6H), 2.23-0.81 (m, 27H), 1.69 (s, 3H), 1.07-0.79 (m, 15H). LC/MS: m/e919.60 (M+H)⁺, 3.27 min (LCMS Method 7).

Step 6. Preparation of benzyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(hydroxymethyl)cyclohex-3-ene-1-carboxylate

To a solution of(1-((benzyloxy)carbonyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-en-1-yl)methylbenzoate (1.0 g, 1.09 mmol) in MeOH (15 mL) was added 1N NaOH (1.09 mL,1.09 mmol). The mixture was stirred at RT for 12 h, neutralized withsaturated aqueous citric acid and the solvent was removed in vacuo. Theresidue was dissolved in EtOAc, washed with brine, dried over Na₂SO₄,and concentrated in vacuo to give the desired product (56% yield withtrace amount methyl ester by product) without further purification.LC/MS: m/e 815 (M+H)⁺, 4.803 min (LCMS Method 7). For methyl ester:LC/MS: m/e 739.55 (M+H)⁺, 4.615 min (LCMS Method 7).

Step 7. Preparation of benzyl1-((aryloxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylate

To a solution of benzyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(hydroxymethyl)cyclohex-3-ene-1-carboxylate(1 eq) in DMF at −78° C. was added KOtBu (2 eq). The resulted mixturewas stirred for 20 minutes before the addition of Ar—X (2 eq). Then thereaction was warmed to RT and stirred overnight. The reaction mixturewas diluted with EtOAc, washed with water, dried over Na₂SO₄, andconcentrated in vacuo to give crude product which was either used innext step without further purification or purified by silica gelchromatography with ethyl acetate/hexanes as eluents.

Step 8. Preparation of tert-butyldimethylsilyl1-((aryloxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylate

To a solution of the crude product (1 eq) from general procedure F, step7 in DCE (3 mL) was added TEA (1.6 eq), 1-Butyldimethylsilane (2.0 eq),and palladium acetate (0.25 eq). The mixture was flushed with N₂ for 5minutes and then heated at 60° C. for 2-6 hours. The reaction mixturewas cooled to room temperature and was filtered through a pad of celiteand silica gel and washed with 50% EtOAc in hexanes, then withdichloromethane. The filtrate was concentrated under reduced pressureand the crude product obtained was used in the next step withoutadditional purification.

Step 9. Preparation of1-((aryloxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylicacid

To a solution of the crude product (1 eq) from general procedure F, step8 in THF (3 mL) was added a solution of TBAF (1.6 eq) in THF. Theresulting mixture was stirred for 2 hours. The solution was purified byreverse phase preparative HPLC. Fractions containing the desired productwere collected and dried to afford the desired1-((aryloxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylicacid.

Example 37 Preparation of1-(((4-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylicacid

Step 1-6: General Procedure F Steps 1-6 Step 7. Preparation of benzyl1-(((4-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 22% yield as a solid, following theprocedure described in general procedure F, step 7, using2-fluoroisonicotinonitrile as the reactant. LC/MS m/z M+1=917.65, 4.765min (LCMS Method 7).

Step 8. Preparation of tert-butyldimethylsilyl1-(((4-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylate

The title compound was prepared as a solid, following the proceduredescribed in general procedure F, step 8, using benzyl1-(((4-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylateas the reactant. LC/MS m/z M+1=941.75, 3.467 min (LCMS Method 7).

Step 9.1-(((4-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylicacid was prepared in 10.7% yield as a solid, following the proceduredescribed in general procedure F, step 9, using tert-butyldimethylsilyl1-(((4-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylateas the reactant. LC/MS: m/e 827.60 (M+H)⁺, 3.00 min (LCMS Method 7). ¹HNMR (400 MHz, CHLOROFORM-d) δ 8.31 (d, J=5.0 Hz, 1H), 7.21 (dd, J=5.1,1.1 Hz, 1H), 7.14 (s, 1H), 5.36 (br. s., 1H), 5.21 (d, J=4.8 Hz, 1H),4.84 (s, 1H), 4.75 (s, 1H), 4.52 (dd, J=4.3, 10.3 Hz, 1H), 4.45 (dd,J=1.8, 10.3 Hz, 1H), 3.28-3.15 (m, 8H), 3.14-3.06 (m, 4H), 2.98-2.87 (m,1H), 2.76 (td, J=11.1, 5.4 Hz, 1H), 2.45-2.58 (m, 1H), 2.35-2.21 (d,J=8.5 Hz, 1H), 2.21-1.98 (m, 8H), 1.92-1.73 (m, 2H), 1.77 (s, 3H),1.73-1.41 (m, 11H), 1.40-1.28 (m, 2H), 1.27-1.08 (m, 2H), 1.18 (s, 3H),1.11 (s, 3H), 1.00-0.99 (m, 3H), 0.96 (s, 3H), 0.93 (s, 3H).

Example 38 Preparation of1-(((5-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylicacid

Step 1-6: General Procedure F Steps 1-6 Step 7. Preparation of benzyl1-(((5-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylate

The title compound was prepared in 35.5% yield as a solid, following theprocedure described in general procedure F, step 7, using6-fluoronicotinonitrile as the reactant. LC/MS m/z M+1=917.65, 3.136 min(LCMS Method 7).

Step 8: Preparation of tert-butyldimethylsilyl1-(((5-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylate

The title compound was prepared as a solid, following the proceduredescribed in general procedure F, step 8, using benzyl1-(((5-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylateas the reactant. LC/MS m/z M+1=941.70, 3.311 mm (LCMS Method 7).

Step 9.1-(((5-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylicacid was prepared in 10.5% of yield as a solid, following the proceduredescribed in general procedure F, step 9, using tert-butyldimethylsilyl1-(((5-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylateas the reactant. LC/MS: m/e 827.55 (M+H)⁺, 3.049 min (LCMS Method 7). ¹HNMR (400 MHz, METHANOL-d₄) δ 8.52 (d, J=2.3 Hz, 1H), 7.96 (dd, J=8.8,2.3 Hz, 1H), 6.92 (d, J=8.8 Hz, 1H), 5.35 (br. s., 1H), 5.21 (d, J=4.5Hz, 1H), 4.84 (s, 1H), 4.75 (s, 1H), 4.56 (dd, J=2.8, 10.5 Hz, 1H), 4.49(d, J=10.5 Hz, 1H), 3.28-3.15 (m, 8H), 3.14-3.06 (m, 3H), 2.93 (dt,J=14.2, 5.2 Hz, 1H), 2.81-2.71 (m, 1H), 2.67-2.57 (m, 1H), 2.35-2.21 (m,1H), 2.21-1.97 (m, 10H), 1.92-1.72 (m, 5H), 1.77 (s, 3H), 1.72-1.39 (m,11H), 1.39-1.20 (m, 1H), 1.20-1.07 (m, 1H), 1.18 (s, 3H), 1.11 (s, 3H),1.02-0.98 (m, 1H), 0.96 (s, 3H), 0.93 (s, 3H).

Example 39 Preparation of1-(((6-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylicacid

Step 1-6: General Procedure F Steps 1-6 Step 7. Preparation of benzyl1-(((6-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylate

The title compound was prepared as a solid, following the proceduredescribed in general procedure F, step 7, using 6-chloropicolinonitrileas the reactant. LC/MS m/z M+1=917.65, 3.083 min (LCMS Method 7).

Step 8. Preparation of tert-butyldimethylsilyl1-(((6-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylate

The title compound was prepared as a solid, following the proceduredescribed in general procedure F, step 8, using benzyl1-(((6-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylateas the reactant. LC/MS m/z M+1=941.70, 3.516 min (LCMS Method 7).

Step 9.1-(((6-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylicacid was prepared in 17.6% of yield as a solid, following the proceduredescribed in general procedure F, step 9, using tert-butyldimethylsilyl1-(((6-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylateas the reactant. LC/MS: m/e 827.55 (M+H)⁺, 3.003 min (LCMS Method 7). ¹HNMR (400 MHz, METHANOL-d₄) δ 7.84 (dd, J=8.5, 7.3 Hz, 1H), 7.47 (d,J=7.3 Hz, 1H), 7.09 (d, J=8.5 Hz, 1H), 5.38 (br. s., 1H), 5.24 (d, J=6.0Hz, 1H), 4.86 (s, 1H), 4.76 (s, 1H), 4.59-4.53 (m, 1H), 4.48-4.43 (m,1H), 3.30-3.17 (m, 8H), 3.09-3.17 (m, 3H), 2.98-2.89 (m, 1H), 2.78 (td,J=10.9, 5.5 Hz, 1H), 2.65 (br. d, J=15.8 Hz, 1H), 2.37-2.00 (m, 9H),1.94-1.74 (m, 5H), 1.79 (s, 3H), 1.74-1.44 (m, 11H), 1.44-1.29 (m, 2H),1.26-1.10 (m, 2H), 1.20 (s, 3H), 1.13 (s, 3H), 1.03-1.02 (m, 3H),1.00-0.99 (m, 3H), 0.95 (s, 3H).

Example 40 Preparation of1-(((6-carbamoylpyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylicacid

Step 1. Preparation of methyl1-(((6-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylate

The title compound was prepared as a solid, following the proceduredescribed in general procedure F, step 7, using 6-chloropicolinonitrileand methyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(hydroxymethyl)cyclohex-3-ene-1-carboxylateinstead of benzyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(hydroxymethyl)cyclohex-3-ene-1-carboxylateas the reactants. LC/MS m/z M+1=841.60, 3.164 min (LCMS Method 7).

Step 2.1-(((6-carbamoylpyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylicacid was prepared in a yield of 19.9% as a solid, following theprocedure described in general procedure E, step 4, using methyl1-(((6-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylateas the reactant. LC/MS: m/e 845.60 (M+H)⁺, 2.931 min (LCMS Method 7). ¹HNMR (400 MHz, METHANOL-d₄) δ 7.84 (dd, J=8.3, 7.3 Hz, 1H), 7.72 (dd,J=73, 0.8 Hz, 1H), 6.99 (d, J=7.8 Hz, 1H), 5.39 (br. s., 1H), 5.23 (d,J=4.3 Hz, 1H), 4.87 (s, 1H), 4.77 (s, 1H), 4.61-4.56 (m, 1H), 4.55-4.50(m, 1H), 3.25 (d, J=8.8 Hz, 5H), 3.20 (br. s., 2H), 3.17-3.09 (m, 3H),2.97-2.87 (m, 1H), 2.77 (d, J=5.3 Hz, 1H), 2.68 (d, J=13.6 Hz, 1H),2.41-1.99 (m, 9H), 1.94-1.68 (m, 6H), 1.79 (s, 3H), 1.68-1.44 (m, 9H),1.43-1.30 (m, 3H), 1.29-1.11 (m, 2H), 1.19 (s, 3H), 1.13 (s, 3H), 1.01(s, 3H), 0.98 (s, 3H), 0.95 (s, 3H).

Preparation of 4-(methylsulfonyl)cyclohexanone

To a solution of (methylsulfonyl)ethene (10.0 g, 94 mmol) in benzene (50mL) was added (buta-1,3-dien-2-yloxy)trimethylsilane (14.07 g, 99 mmol)and hydroquinone (20 mg, 0.182 mmol). The mixture was degassed severaltimes at −78° C. prior to heating. The contents were sealed and heatedat 105° C. for 48 hours. The reaction was analyzed by NMR in CDCl3 thatshowed about 10% of the vinyl sulfone residue. Additional(buta-1,3-dien-2-yloxy)trimethylsilane (4 mL) was added and heatingresumed for another 48 hours. NMR analysis again at 72 hrs time pointshowed further reduction of the amount of vinyl sulfone (3%). The samplefrom the NMR tube was combined the reaction mixture and evaporated to athick gum under vacuum at room temperature (19° C.). The mixture wasrediluted with acetone (250 mL) resulting in the formation of a clearsolution. The mixture was chilled in an ice bath until cold. 4 mL of0.25 N HCl (pre-chilled in the same ice-bath) was added resulting in theformation a cloudy mixture, which became clear after 15 minutes ofstirring at 0° C., and then returned to a cloudy state in another 10minutes, it remained turbid for the rest of stirring period. A 50 μLaliquot was removed, flash dried into a film, and was analyzed by NMR inCDCl₃. NMR showed 7% of vinyl sulfone relative to the desired product.The acetone solution was filtered through a short bed of silica geltype-H after a total reaction time of about one hour, and was thenwashed with more acetone. The filtrate was concentrated on the rotovaporat 19° C. bath temperature. The crude product was sub-divided into twoparts, 7.75 gm each, for purification. The product was purified bycolumn chromatography on silica gel (30% ethyl acetate→100% ethylacetate in hexanes; two 330 g columns) to afford4-(methylsulfonyl)cyclohexanone (16.7 g, 100% yield) as a white solid:¹H NMR (400 MHz, CHLOROFORM-d) δ 3.29 (tt, J=11.0, 3.9 Hz, 1H), 2.94 (s,3H), 2.73-2.62 (m, 2H), 2.58-2.37 (m, 4H), 2.15 (qd, J=11.9, 4.5 Hz,2H).

Preparation of2-(cis-1-hydroxy-4-(methylsulfonyl)cyclohexyl)acetaldehyde and2-(trans-1-hydroxy-4-(methylsulfonyl)cyclohexyl)acetaldehyde

Step 1. Preparation of (cis)-1-allyl-4-(methylsulfonyl)cyclohexanol and(trans)-1-allyl-4-(methylsulfonyl)cyclohexanol

To a solution of 4-(methylsulfonyl)cyclohexanone (1.03 g, 5.84 mmol) inTHF (40 mL) at 0° C. was added via cannula allylmagnesium bromide (7.60mL, 7.60 mmol). The reaction mixture was stirred at 0° C. for 30 min.The reaction was quenched by the addition of saturated NH₄Cl solution(25 mL). The mixture was transferred to a separatory funnel and theaqueous layer was extracted with ethyl acetate (5×50 mL). The combinedorganic layers were washed with brine (20 mL), dried over MgSO₄,filtered, and concentrated. The product was purified by columnchromatography on silica gel (70% ethyl acetate with 1% methanol/30%hexanes→100% ethyl acetate with 1% methanol; 40 g column) to afford(cis)-1-allyl-4-(methylsulfonyl)cyclohexanol (374 mg, 1.713 mmol, 29%yield) as a white solid and(trans)-1-allyl-4-(methylsulfonyl)cyclohexanol (551 mg, 2.52 mmol, 43%yield) as a colorless oil.

(cis)-1-allyl-4-(methylsulfonyl)cyclohexanol

¹H NMR (400 MHz, CDCl₃) δ 5.96-5.79 (m, 1H), 5.26-5.21 (m, 1H), 5.18(ddt, J=17.1, 2.1, 1.2 Hz, 1H), 2.85 (s, 3H), 2.80 (tt, J=12.5, 3.6 Hz,1H), 2.25 (d, J=7.5 Hz, 2H), 2.15-2.07 (m, 2H), 1.97 (qd, J=13.0, 3.8Hz, 2H), 1.88-1.81 (m, 2H), 1.52-1.42 (m, 2H); ¹³C NMR (100 MHz, CDCl₃)δ 132.50, 120.02, 69.06, 62.26, 47.86, 36.85, 35.67, 21.13. Thestructure of (cis)-1-allyl-4-(methylsulfonyl)cyclohexanol was confirmedby X-ray crystallography.

(trans)-1-allyl-4-(methylsulfonyl)cyclohexanol

¹H NMR (400 MHz, CDCl₃) δ 5.88 (ddt, J=17.2, 10.1, 7.4 Hz, 1H),5.28-5.16 (m, 2H), 2.98-2.91 (m, 1H), 2.90 (s, 3H), 2.35 (d, J=7.5 Hz,2H), 2.23-2.14 (m, 2H), 2.02-1.93 (m, 2H), 1.90-1.78 (m, 2H), 1.57-1.46(m, 2H); ¹³C NMR (100 MHz, CDCl₃) δ 132.62, 120.19, 69.20, 62.41, 48.00,36.98, 35.83, 21.29.

Step 2a. Preparation of2-((cis)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)acetaldehyde

(trans)-1-Allyl-4-(methylsulfonyl)cyclohexanol (3.4 g, 15.57 mmol) wasdissolved in CH₂Cl₂ (160 mL) and MeOH (32.0 mL) in a 500 mL round bottomflask. N-Methylmorpholine-N-oxide (NMO) (2.189 g, 18.69 mmol) was addedand the mixture was cooled to −78° C. [Schwartz, C., Raible, J., Mott,K., Dussault, P. H. Org. Lett. 2006, 8, 3199-3201], Ozone was bubbledthrough the reaction mixture until the solution was saturated with ozone(turned into a blue color) and several minutes thereafter (total time 25min). Nitrogen was then bubbled through the reaction mixture until thedisappearance of the blue color. Dimethyl sulfide (11.52 mL, 156 mmol)was then added and the reaction mixture was stirred at 0° C. for 16 h.The mixture was concentrated under vacuum. The product was purified bycolumn chromatography on silica gel (50% ethyl acetate with 1%methanol/50% hexanes→95% ethyl acetate with 1% methanol/5% hexanes; 330g column) to afford2-((1s,4s)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)acetaldehyde (3.31 g,15.03 mmol, 96% yield) as a white solid: ¹H NMR (400 MHz, CHLOROFORM-d)δ 9.87 (t, J=1.1 Hz, 1H), 2.85 (s, 3H), 2.82-2.76 (m, 1H), 2.67 (d,J=1.3 Hz, 2H), 2.13-1.98 (m, 6H), 1.50-1.38 (m, 2H); ¹³C NMR (101 MHz,CHLOROFORM-d) δ 202.5, 68.9, 61.9, 54.9, 36.8, 35.9, 20.8.

Step 2b. Preparation of2-((trans)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)acetaldehyde

(1r,4r)-1-Allyl-4-(methylsulfonyl)cyclohexanol (2 g, 9.16 mmol) wasdissolved in CH₂Cl₂ (80 mL) and MeOH (16.00 mL) in a 500 mL round bottomflask. N-Methylmorpholine-N-oxide (NMO) (1.288 g, 10.99 mmol) was addedand the mixture was cooled to −78° C. [Schwartz, C., Raible, J., Mott,K., Dussault, P. H. Org. Lett. 2006, 8, 3199-3201], Ozone (excess) wasbubbled through the reaction mixture until the solution was saturatedwith ozone (turned into a blue color) and several minutes thereafter(total time 25 min). Nitrogen was then bubbled through the reactionmixture until the disappearance of the blue color. Dimethyl sulfide(6.78 mL, 92 mmol) was then added and the reaction mixture was stirredat 0° C. for 16 h. The mixture was concentrated under vacuum. Theproduct was purified by column chromatography on silica gel (70% ethylacetate with 5% methanol/30% hexanes→100% ethyl acetate with 5%methanol; 220 g column) to afford2-((1r,4r)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)acetaldehyde (1.58 g,7.17 mmol, 78% yield) as a white solid: ¹H NMR (400 MHz, CHLOROFORM-d) δ9.82 (t, J=1.8 Hz, 1H), 2.99-2.88 (m, 1H), 2.85 (s, 3H), 2.67 (d, J=1.8Hz, 2H), 2.20-2.10 (m, 2H), 2.06-1.98 (m, 2H), 1.74 (dtd, J=14.0, 10.6,3.5 Hz, 2H), 1.61-1.50 (m, 2H); ¹³C NMR (101 MHz, CHLOROFORM-d) δ 202.4,70.0, 59.3, 50.3, 38.2, 34.9, 21.1.

Example A1. Preparation of4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1s,4R)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-2-yloxy)methyl)cyclohex-3-enecarboxylicacid

Step 1. Preparation of ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1s,4R)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-2-yloxy)methyl)cyclohex-3-enecarboxylate

A mixture of ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-2-yloxy)methyl)cyclohex-3-enecarboxylate(65 mg, 0.097 mmol),2-((1s,4s)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)acetaldehyde (47.1 mg,0.214 mmol), and borane-2-picoline complex (22.86 mg, 0.214 mmol) inMeOH (1 mL) and acetic acid (0.2 mL) was stirred at room temperature for16 h. The mixture was transferred to a separatory funnel containingsaturated aqueous sodium bicarbonate solution (10 mL) and saturatedaqueous sodium carbonate solution (2 mL). The aqueous layer wasextracted with dichloromethane (4×20 mL). The combined organic layerswere washed with brine (10 mL), dried over MgSO₄, filtered, andconcentrated. The product was purified by column chromatography onsilica gel (10% 9:1 acetone:methanol/90% hexanes→65% 9:1acetone:methanol/35% hexanes; 24 g column, λ=220 nm) to afford ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1s,4R)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-2-yloxy)methyl)cyclohex-3-enecarboxylate(69 mg, 81% yield) as a colorless solid: ¹H NMR (500 MHz, CHLOROFORM-d)δ 8.15 (dd, J=5.0, 1.8 Hz, 1H), 7.56 (ddd, J=8.5, 6.9, 2.0 Hz, 1H), 6.87(td, J=6.1, 0.7 Hz, 1H), 6.73 (d, J=8.4 Hz, 1H), 5.37 (br. s., 1H), 5.20(d, J=6.1 Hz, 1H), 4.75 (d, J=1.5 Hz, 1H), 4.62 (s, 1H), 4.50-4.44 (m,1H), 4.43-4.37 (m, 1H), 4.21-4.10 (m, 2H), 2.85 (s, 3H), 2.84-2.67 (m,4H), 2.55 (td, J=10.8, 5.5 Hz, 1H), 2.22-0.88 (m, 43H), 1.70 (s, 3H),1.21 (t, J=7.1 Hz, 3H), 1.06 (s, 3H), 0.98 (s, 3H), 0.87 (s, 3H); LC/MSm/e 873.7 [(M+H)⁺, calcd for C₅₃H₈₁N₂O₆S 873.6], t_(R)=4.67 min (LCMSMethod 14).

Step 2. A solution of ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1s,4R)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-2-yloxy)methyl)cyclohex-3-enecarboxylate(65 mg, 0.074 mmol) in dioxane (1 mL) and MeOH (0.5 mL) was treated withsodium hydroxide (0.372 mL, 0.744 mmol, 2M aq). The reaction mixture washeated at 50° C. for 3 h and then at 60° C. for 6 h. The mixture wascooled to room temperature, and was partially neutralized by theaddition of 2 N HCl (200 uL). The mixture was filtered through a syringefilter, and was purified by reverse phase HPLC (Preparative HPLC Method1). The product (61.7 mg) contained an impurity (ca. 6%). The productwas repurified by reverse phase HPLC (Preparative HPLC Method 2). Theorganic solvent was evaporated on the rotovapor and the aqueous mixturewas lyophilized to afford4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1s,4R)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-2-yloxy)methyl)cyclohex-3-enecarboxylicacid, TFA (48.4 mg, 67% yield) as a white amorphous solid: ¹H NMR (500MHz, Acetic Acid-d₄) δ 8.29 (dd, J=5.3, 1.7 Hz, 1H), 7.90-7.82 (m, 1H),7.13-7.08 (m, 1H), 6.99 (d, J=8.4 Hz, 1H), 5.41 (br. s., 1H), 5.26 (d,J=5.8 Hz, 1H), 4.83 (s, 1H), 4.73 (s, 1H), 4.56-4.50 (m, 1H), 4.49-4.44(m, 1H), 3.48-3.34 (m, 2H), 3.09-2.99 (m, 1H), 2.96 (s, 3H), 2.89-2.79(m, 1H), 2.72 (d, J=16.0 Hz, 1H), 2.32-1.32 (m, 35H), 1.75 (s, 3H), 1.17(s, 3H), 1.13 (d, J=7.5 Hz, 2H), 1.09 (s, 3H), 1.02 (d, J=3.7 Hz, 3H),0.99 (d, J=3.7 Hz, 3H), 0.95 (s, 3H); LC/MS m/e 845.6 [(M+H)¹, calcd forC₅₁H₇₇N₂O₆S 845.6], t_(R)=4.36 min (LCMS Method 14); HPLC (AnalyticalHPLC Method 1): t_(R)=18.86 min; HPLC (Analytical HPLC Method 2):(R=20.24 min.

Example A2. Preparation of4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1r,4S)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-2-yloxy)methyl)cyclohex-3-enecarboxylicacid

Step 1. Preparation of ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1r,4S)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-2-yloxy)methyl)cyclohex-3-enecarboxylate

A mixture of ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-2-yloxy)methyl)cyclohex-3-enecarboxylate(65 mg, 0.097 mmol),2-((1r,4r)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)acetaldehyde (47.1 mg,0.214 mmol), and borane-2-picoline complex (22.86 mg, 0.214 mmol) inMeOH (1 mL) and acetic acid (0.2 mL) was stirred at room temperature for16 h. The reaction was not complete. Additional2-((1r,4r)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)acetaldehyde (21 mg,0.097 mmol, 1 eq) was then added and 1 h later borane-2-picoline complex(10 mg, 0.097 mmol, 1 eq) was added to the reaction mixture and themixture was stirred at room temperature for 3 h. The mixture wastransferred to a separatory funnel containing saturated aqueous sodiumbicarbonate solution (10 mL) and saturated aqueous sodium carbonatesolution (2 mL). The aqueous layer was extracted with dichloromethane(4×20 mL). The combined organic layers were washed with brine (10 mL),dried over MgSO₄, filtered, and concentrated. The product was purifiedby column chromatography on silica gel (10% 9:1 acetone:methanol/90%hexanes→65% 9:1 acetone:methanol/35% hexanes; 24 g column, λ=220 nm) toafford ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1r,4S)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-2-yloxy)methyl)cyclohex-3-enecarboxylate(42.4 mg, 50% yield) as a colorless foam: ¹H NMR (500 MHz, CHLOROFORM-d)δ 8.14 (dd, J=5.0, 1.8 Hz, 1H), 7.61-7.54 (m, 1H), 6.87 (ddd, J=7.0,5.1, 0.8 Hz, 1H), 6.73 (d, J=8.4 Hz, 1H), 5.36 (br. s., 1H), 5.20 (d,J=6.0 Hz, 1H), 4.72 (d, J=1.4 Hz, 1H), 4.60 (s, 1H), 4.51-4.44 (m, 1H),4.43-4.36 (m, 1H), 4.21-4.10 (m, 2H), 2.99-2.91 (m, 1H), 2.89 (s, 3H),2.83-2.76 (m, J=12.1 Hz, 1H), 2.72-2.62 (m, 2H), 2.59-2.51 (m, 1H),2.21-0.88 (m, 43H), 1.69 (s, 3H), 1.21 (t, J=7.1 Hz, 3H), 1.06 (s, 3H),0.98 (s, 3H), 0.87 (s, 3H); LC/MS m/e 873.7 [(M+H)⁺, calcd forC₅₃H₈₁N₂O₆S 873.6], t_(R)=4.62 min (LCMS Method 14).

Step 2. A solution of ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1r,4S)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-2-yloxy)methyl)cyclohex-3-enecarboxylate(42 mg, 0.048 mmol) in dioxane (1 mL) and MeOH (0.5 mL) was treated withsodium hydroxide (0.361 mL, 0.721 mmol, 2 M aq). The reaction mixturewas heated at 60° C. for 24 h. Additional sodium hydroxide (0.120 mL,0.240 mmol, 5 eq, 2 M aq) was added and the reaction mixture was heatedat 70° C. for 8 h. The reaction was complete. The mixture was cooled toroom temperature, and was partially neutralized by the addition of 2 NHCl (400 uL). The mixture was filtered through a syringe filter, and waspurified by reverse phase HPLC (Preparative HPLC Method 3). The organicsolvent was evaporated on the rotovapor and the aqueous mixture waslyophilized to afford 4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1r,4S)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-((pyridin-2-yloxy)methyl)cyclohex-3-enecarboxylicacid, TFA (31.3 mg, 67% yield) as a white amorphous solid. ¹H NMR (500MHz, Acetic Acid-d₄) δ 8.30-8.25 (m, 1H), 7.85-7.78 (m, 1H), 7.10-7.04(m, 1H), 6.95 (dd, J=8.5, 0.6 Hz, 1H), 5.41 (br. s., 1H), 5.26 (d, J=6.0Hz, 1H), 4.83 (s, 1H), 4.72 (s, 1H), 4.55-4.49 (m, 1H), 4.48-4.43 (m,1H), 3.46-3.37 (m, 1H), 3.36-3.28 (m, 1H), 3.18-3.10 (m, 1H), 2.98 (s,3H), 2.91-2.81 (m, 1H), 2.71 (d, J=16.3 Hz, 1H), 2.32-1.32 (m, 35H),1.75 (s, 3H), 1.16-1.12 (m, 2H), 1.14 (s, 3H), 1.09 (s, 3H), 1.02 (d,J=3.7 Hz, 3H), 0.99 (d, J=3.2 Hz, 3H), 0.94 (s, 3H); LC/MS m/e 845.6[(M+H)⁺, calcd for C₅₁H₇₇N₂O₆S 845.6], t_(R)=4.33 min (LCMS Method 14);HPLC (Analytical HPLC Method 1): t_(R)=18.86 min; HPLC (Analytical HPLCMethod 2): t_(R)=20.48 min.

Example A3. Preparation of(S)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1s,4R)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid

Step 1. Preparation of((S)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(ethoxycarbonyl)cyclohex-3-en-1-yl)methylbenzoate

To a flask containing(1R,3aS,5aR,5bR,7aR,11aR,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yltrifluoromethanesulfonate (1.00 g, 1.79 mmol) was added(R)-(1-(ethoxycarbonyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-en-1-yl)methylbenzoate (1.337 g, 3.23 mmol), potassium phosphate tribasic (1.52 g,7.17 mmol), 2-dicyclohexylphosphino-2′,6′-dimethoxybiphenyl (S-Phos)(0.055 g, 0.134 mmol) and palladium(II) acetate (0.020 g, 0.090 mmol).The mixture was diluted with 1,4-dioxane (25 mL) and water (6.25 mL),then was flushed with N₂ and heated at 75° C. for 16 h. The mixture wascooled to rt. The mixture was diluted with water (100 mL) and extractedwith ethyl acetate (3×75 mL). The organic layers were washed with brine(150 mL), dried over magnesium sulfate, filtered and concentrated underreduced pressure. The residue was purified by column chromatography onsilica gel (50% ethyl acetate with 4% MeOH and 0.8% aminoniumhydroxide/50% hexanes→70% ethyl acetate with 4% MeOH and 0.8% aminoniumhydroxide/30% hexanes, 120 g column) to afford((S)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(ethoxycarbonyl)cyclohex-3-en-1-yl)methylbenzoate (1.15 g, 92% yield) as an off-white solid: ¹H NMR (400 MHz,CHLOROFORM-d) δ 8.06-8.00 (m, 2H), 7.62-7.55 (m, 1H), 7.49-7.41 (m, 2H),5.38 (br. s., 1H), 5.22 (dd, J=6.3, 1.8 Hz, 1H), 4.75 (d, J=2.0 Hz, 1H),4.62 (dd, J=2.3, 1.3 Hz, 1H), 4.52-4.40 (m, 2H), 4.20 (qd, J=7.2, 2.1Hz, 2H), 2.70 (d, J=18.3 Hz, 1H), 2.56 (td, J=10.9, 5.3 Hz, 1H),2.35-1.95 (m, 6H), 1.91-1.81 (m, 1H), 1.78-1.13 (m, 20H), 1.71 (s, 3H),1.24 (t, J=7.3 Hz, 3H), 1.09 (s, 3H), 1.00 (s, 3H), 0.98 (s, 3H), 0.93(s, 3H), 0.88 (s, 3H); LC/MS m/e 696.7 [(M+H)⁺, calcd for C₄₆H₆₅NO₄696.5], t_(R)=2.60 min (LCMS Method 15).

Step 2. Preparation of (S)-ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(hydroxymethyl)cyclohex-3-enecarboxylate

To a solution of((S)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(ethoxycarbonyl)cyclohex-3-en-1-yl)methylbenzoate (1.07 g, 1.537 mmol) in THF (10 mL) and MeOH (1 mL) was addedsodium hydroxide (1.691 mL, 1.691 mmol). The reaction mixture wasstirred at r.t. for 14 h. The solid was removed by filtration. Themixture was transferred to a separatory funnel containing saturatedaqueous NaHCO₃ solution (10 mL)/water (10 mL). The aqueous layer wasextracted with 5% methanol in ethyl acetate (5×25 mL). The combinedorganic layers were washed with brine (10 mL). The brine wash wasreextracted with 5% methanol in ethyl acetate. The combined organiclayers were dried over MgSO₄, filtered, and concentrated to afford(S)-ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(hydroxymethyl)cyclohex-3-enecarboxylate(0.535 g, 59% yield) as a white solid. The crude product was useddirectly in the next step. ¹H NMR (400 MHz, CHLOROFORM-d) δ 5.34 (t,J=3.8 Hz, 1H), 5.20 (dd, J=6.1, 1.9 Hz, 1H), 4.75 (d, J=2.0 Hz, 1H),4.63 (d, J=1.3 Hz, 1H), 4.21 (q, J=7.0 Hz, 2H), 3.70 (s, 2H), 2.62-2.51(m, 2H), 2.23-2.15 (m, 2H), 2.09-1.92 (m, 4H), 1.83-1.12 (m, 21H), 1.72(s, 3H), 1.30 (t, J=1.2 Hz, 3H), 1.09 (s, 3H), 0.99 (s, 3H), 0.98 (s,3H), 0.93 (s, 3H), 0.88 (s, 3H); LC/MS (ESI) mte 614.6 [(M+H)⁺, calcdfor C₃₉H₆₁NO₃Na 614.5], =4.28 min (LCMS Method 14).

Step 3. Preparation of (S)-ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((3-cyanopyridin-2-yl)oxy)methyl)cyclohex-3-enecarboxylate

To a solution of (S)-ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(hydroxymethyl)cyclohex-3-enecarboxylate(495 mg, 0.836 mmol) and 2-fluoronicotinonitrile (204 mg, 1.673 mmol) inTHF (7 mL) and DMF (1 mL) at 0° C. was added potassium tert-butoxide(1.004 mL, 1.004 mmol). The cooling bath was removed and the reactionmixture was stirred at 20° C. for 1.5 h. The mixture was transferred toa separatory funnel containing saturated aqueous NaHCO₃ solution (15mL). The aqueous layer was extracted with ethyl acetate (4×25 mL). Thecombined organic layers were washed with brine (15 mL), dried overMgSO₄, filtered, and concentrated. The product was purified by columnchromatography on silica gel (50% of a 5% methanol in ethyl acetatesolution/50% hexanes→100% of a 5% methanol in ethyl acetate solution; 40g column) to afford (S)-ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((3-cyanopyridin-2-yl)oxy)methyl)cyclohex-3-enecarboxylate(344 mg, 59% yield) as an off-white solid: ¹H NMR (400 MHz,CHLOROFORM-d) δ 8.34 (dd, J=5.0, 2.0 Hz, 1H), 7.88 (dd, J=7.5, 2.0 Hz,1H), 6.99 (dd, J=7.5, 5.0 Hz, 1H), 5.38 (br. s., 1H), 5.21 (dd, J=6.1,1.6 Hz, 1H), 4.75 (d, J=2.3 Hz, 1H), 4.62 (dd, J=2.1, 1.4 Hz, 1H), 4.57(s, 2H), 4.25-4.15 (m, 2H), 2.78-2.68 (m, 1H), 2.56 (td, J=10.9, 5.1 Hz,1H), 2.35-1.89 (m, 6H), 1.79-1.11 (m, 21H), 1.71 (s, 3H), 1.27 (t, J=6.8Hz, 3H), 1.09 (s, 3H), 0.99 (s, 3H), 0.98 (s, 3H), 0.93 (s, 3H), 0.88(s, 3H); LC/MS (ESI) m/e 694.7 [(M+H)⁺, calcd for C₄₅H₆₄N₃O₃ 694.5],=4.52 min (LCMS Method 14).

Step 4. Preparation of (S)-ethyl1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1s,4R)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylate

(S)-Ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((3-cyanopyridin-2-yl)oxy)methyl)cyclohex-3-enecarboxylate(150 mg, 0.216 mmol) and 2-((1s,4s)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)acetaldehyde (76 mg, 0.346mmol) were dissolved in MeOH (1.6 mL) and acetic acid (0.32 mL).Borane-2-picoline complex (37.0 mg, 0.346 mmol) was added and themixture was stirred at room temperature for 14 h. The mixture wastransferred to a separatory funnel containing saturated aqueous sodiumbicarbonate solution (3 mL) and sodium carbonate solution (2 mL). Theaqueous layer was extracted with ethyl acetate (5×10 mL). The combinedorganic layers were washed with brine (5 mL), dried over MgSO₄,filtered, and concentrated. The product was purified by columnchromatography on silica gel (30% ethyl acetate with 5% methanol/70%hexanes→100% ethyl acetate with 5% methanol; 24 g column, 25 mingradient) to afford (S)-ethyl1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1s,4R)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylate(134.6 mg, 69% yield) as a white foam: ¹H NMR (500 MHz, CHLOROFORM-d) δ8.34 (dd, J=5.0, 2.0 Hz, 1H), 7.88 (dd, J=7.6, 1.9 Hz, 1H), 6.99 (dd,J=7.5, 5.0 Hz, 1H), 5.37 (br. s., 1H), 5.20 (dd, J=6.2, 1.6 Hz, 1H),4.74 (d, J=1.5 Hz, 1H), 4.61 (s, 1H), 4.56 (s, 2H), 4.24-4.15 (m, 2H),2.85 (s, 3H), 2.83-2.67 (m, 4H), 2.55 (td, J=10.9, 5.6 Hz, 1H),2.31-0.88 (m, 37H), 1.70 (s, 3H), 1.26 (t, J=7.1 Hz, 3H), 1.05 (s, 3H),0.98 (s, 3H), 0.98 (s, 3H), 0.93 (s, 3H), 0.87 (s, 3H); LC/MS (ESI) m/e898.7 [(M+H)⁺, calcd for C₅₄H₈₀N₃O₆S 898.6], =4.44 min (LCMS Method 14).

Step 5. To a solution of (S)-ethyl1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1s,4R)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylate(123 mg, 0.137 mmol) in dioxane (4 mL) and MeOH (2 mL) was added lithiumhydroxide (2 mL, 2.00 mmol, 1 M aq). The mixture was heated at 60° C.for 12.5 h. Only a small amount of starting material was detected byLC/MS (LCMS Method 16). The reaction was stopped at this point due tocompeting hydrolysis of the nitrile group to the corresponding amide.The mixture was cooled to room temperature and was partially neutralizedby the addition of 6 N HCl (250 μL). The mixture was then filteredthrough a syringe filter, and was purified by reverse phase HPLC (5injections) (Preparative HPLC Method 4). The organic solvent wasevaporated on the rotovapor and the aqueous mixture was lyophilized toafford(S)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1s,4R)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid, TFA (51.6 mg, 38% yield) as a white amorphous solid: ¹H NMR (500MHz, Acetic Acid-d₄) δ 8.42 (dd, J=5.1, 1.9 Hz, 1H), 8.05 (dd, J=7.6,1.9 Hz, 1H), 7.11 (dd, J=7.6, 5.1 Hz, 1H), 5.43 (s, 1H), 5.27 (d, J=4.7Hz, 1H), 4.83 (s, 1H), 4.72 (s, 1H), 4.68-4.61 (m, 2H), 3.47-3.33 (m,2H), 3.08-2.99 (m, 1H), 2.96 (s, 3H), 2.90-2.81 (m, 1H), 2.74 (d, J=15.6Hz, 1H), 2.38-1.13 (m, 37H), 1.75 (s, 3H), 1.18 (s, 3H), 1.09 (s, 3H),1.03 (s, 3H), 0.99 (s, 3H), 0.95 (s, 3H); LC/MS (ESI) m/e 870.6 [(M+H)⁺,calcd for C₅₂H₇₆N₃O₆S 870.5], t_(R)=1.31 min (LCMS Method 16); HPLC(Analytical HPLC Method 3): t_(R)=12.19 min; HPLC (Analytical HPLCMethod 4): t_(R)=11.64 min.

Alternate Route for the Preparation of Example A3

Preparation of(S)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1s,4R)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid

Step 1. Preparation of((S)-1-(ethoxycarbonyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1s,4R)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-en-1-yl)methylbenzoate

((S)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(ethoxycarbonyl)cyclohex-3-en-1-yl)methyl benzoate (7.63 g, 10.96 mmol), and2-((1s,4s)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)acetaldehyde (3.86 g,17.54 mmol) were dissolved in MeOH (30 mL) and acetic acid (6 mL).Borane-2-picoline complex (1.876 g, 17.54 mmol) was added and themixture was stirred at room temperature for 14 h. The mixture wastransferred to a separatory funnel containing saturated aqueous sodiumbicarbonate solution (50 mL) and sodium carbonate solution (50 mL). Theaqueous layer was extracted with ethyl acetate (7×100 mL). The combinedorganic layers were washed with brine (25 mL), dried over MgSO₄,filtered, and concentrated. The product was purified by columnchromatography on silica gel (30% ethyl acetate with 5% methanol/70%hexanes→100% ethyl acetate with 5% methanol; 330 g column, 30 mingradient) to afford((S)-1-(ethoxycarbonyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1s,4R)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-en-1-yl)methylbenzoate (8.81 g, 89% yield) as a white solid: ¹H NMR (500 MHz,CHLOROFORM-d) δ 8.06-8.00 (m, 2H), 7.61-7.54 (m, 1H), 7.50-7.42 (m, 2H),5.37 (br. s., 1H), 5.21 (dd, J=6.2, 1.6 Hz, 1H), 4.75 (d, J=1.8 Hz, 1H),4.62 (s, 1H), 4.47-4.41 (m, 2H), 4.24-4.16 (m, 2H), 2.85 (s, 3H),2.83-2.65 (m, 4H), 2.55 (td, J=10.9, 5.6 Hz, 1H), 2.33-2.23 (m, 1H),2.20-1.03 (m, 36H), 1.70 (s, 3H), 1.24 (t,7=7.1 Hz, 3H), 1.05 (s, 3H),0.99-0.87 (m, 12H); LC/MS (ESI) m/e 900.4 [(M+H)⁺, calcd for C₅₅H₈₂NO₇S900.6], t_(R)=4.55 min (LCMS Method 14).

Step 2. Preparation of(S)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1s,4R)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(hydroxymethyl)cyclohex-3-enecarboxylicacid

To a solution of((S)-1-(ethoxycarbonyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1s,4R)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-en-1-yl)methylbenzoate (8.00 g, 8.89 mmol) in 1,4-dioxane (160 mL) and methanol (80mL) in a pressure vessel was added lithium hydroxide (89 mL, 89 mmol).The vessel was sealed and the mixture was heated at 65° C. (internaltemperature) for 16 h. The reaction mixture was cooled to roomtemperature and was partially neutralized by the addition of 4 N HCl(15.5 mL, 7 eq). The mixture was then concentrated. The crude productwas taken up in dioxane (40 mL)/methanol (20 mL)/water (5 mL) and wasmade acidic by the addition of TFA (dropwise until acidic). Thesuspension became a solution. The solution contained some suspendedsolid matter. It was passed through a short plug of sand followed byfiltration through a syringe filter. The product was then purified byreverse phase MPLC on a C18 Redi Sep Gold column (150 g) on the biotage(Preparative MPLC Method 1, 6 injections). The organic solvent wasevaporated on the rotovapor and the aqueous mixture was lyophilized toafford(S)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1s,4R)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(hydroxymethyl)cyclohex-3-enecarboxylicacid, TFA (6.57 g, 84% yield) as a white amorphous solid. The productwas then dried further under vacuum in a vacuum dessicator with dryrite.¹H NMR (500 MHz, CHLOROFORM-d) δ 8.54 (br. s., 1H), 8.02 (br. s., 1H),5.34 (br. s., 1H), 5.23-5.16 (m, 1H), 4.78 (s, 1H), 4.70 (s, 1H), 3.76(s, 2H), 3.22 (d, J=3.1 Hz, 2H), 2.86 (s, 3H), 2.83-2.68 (m, 2H), 2.59(d, J=15.3 Hz, 1H), 2.47-2.34 (m, 1H), 2.26-1.06 (m, 36H), 1.71 (s, 3H),1.09 (s, 3H), 1.03 (s, 3H), 0.98 (s, 3H), 0.93 (s, 3H), 0.88 (s, 3H);LC/MS (ESI) m/e 768.4 [(M+H)⁺, calcd for C₄₆H₇₄NO₆S 768.5], (R=3.85 mm.(LCMS Method 14).

Step 3. To a solution of(S)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1s,4R)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(hydroxymethyl)cyclohex-3-enecarboxylicacid, TFA (5.92 g, 6.71 mmol) in THF (80 mL) at 0° C. was added sodiumhydride (2.147 g, 53.7 mmol). The cooling bath was removed and thereaction mixture was stirred at room temperature for 15 min. The mixturewas cooled to 0° C. and 2-fluoronicotinonitrile (3.28 g, 26.8 mmol) inTHF (10 mL) was added via cannula. The reaction mixture was stirred at0° C. for 1.5 h. The reaction was quenched by the addition of aceticacid (3.84 mL, 67.1 mmol, 10 eq). The solution was directly injected ona column and was purified by column chromatography on silica gel (5%methanol in CH₂Cl₂ to elute the high R_(f) material and then 12%methanol in CH₂Cl₂ to elute the product. 6.70 g of product was obtained.The product was then purified further by reverse phase MPLC on a C18Redi Sep Gold column (150 g) on the biotage (Preparative MPLC Method 2,5 injections). The organic solvent was evaporated on the rotovapor andthe aqueous mixture was lyophilized to afford(S)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1s,4R)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid, TFA (5.06 g, 5.14 mmol) as a white amorphous solid.

The product (TFA salt) was then dissolved in MeCN/H₂O (60/40) and wasslowly passed through an AG 1-x2 ion exchange resin chloride form(Bio-Rad 100-200 mesh cat #140-1241, prewashed with 90% acetonitrile/10%water). 140 grams of resin was used. The fractions containing productwere combined and the organic solvent was removed on the rotovapor andwater was frozen and placed on the lyophilizer to afford(S)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1s,4R)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid, HCl (4.26 g, 66% yield) as a white amorphous solid: ¹H NMR (500MHz, Acetic Acid-d₄) δ 8.42 (dd, J=5.1, 1.9 Hz, 1H), 8.05 (dd, J=7.6,1.9 Hz, 1H), 7.11 (dd, J=7.6, 5.1 Hz, 1H), 5.43 (br. s., 1H), 5.27 (d,J=4.6 Hz, 1H), 4.89 (s, 1H), 4.73 (s, 1H), 4.69-4.60 (m, 2H), 3.45-3.33(m, 2H), 3.13 (td, J=10.8, 5.1 Hz, 1H), 3.08-3.00 (m, 1H), 2.97 (s, 3H),2.74 (d, J=15.1 Hz, 1H), 2.61-2.53 (m, 1H), 2.38-1.13 (m, 36H), 1.76 (s,3H), 1.20 (s, 3H), 1.10 (s, 3H), 1.03 (s, 3H), 0.99 (s, 3H), 0.95 (s,3H); LC/MS (ESI) m/e 870.3 [(M+H)⁺, calcd for C₅₂H₇₆N₃O₆S 870.5],t_(R)=4.56 min (LCMS Method 14); HPLC (HPLC Method 3): t_(R)=13.13 min;HPLC (HPLC Method 4): t_(R)=12.46 min.

Example A4. Preparation of(S)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1r,4S)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid

Step 1. Preparation of (S)-ethyl1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1r,4S)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylate

(S)-Ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((3-cyanopyridin-2-yl)oxy)methyl)cyclohex-3-enecarboxylate(150 mg, 0.216 mmol) and2-((1r,4r)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)acetaldehyde (76 mg,0.346 mmol) were dissolved in MeOH (1.6 mL) and acetic acid (0.32 mL).Borane-2-picoline complex (37.0 mg, 0.346 mmol) was added and themixture was stirred at room temperature for 16 h. The mixture wastransferred to a separatory funnel containing saturated aqueous sodiumbicarbonate solution (20 mL). The aqueous layer was extracted with ethylacetate (5×20 mL). The combined organic layers were washed with brine(50 mL), dried over MgSO₄, filtered, and concentrated. The product waspurified by column chromatography on silica gel (30% ethyl acetate with5% methanol/70% hexanes→100% ethyl acetate with 5% methanol; 24 gcolumn) to afford (S)-ethyl1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1r,4S)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylate(131 mg, 68% yield) as a white foam: ¹H NMR (400 MHz, CHLOROFORM-d) δ8.33 (dd, J=5.0, 2.0 Hz, 1H), 7.87 (dd, J=7.5, 2.0 Hz, 1H), 6.98 (dd,J=7,4, 5.1 Hz, 1H), 5.37 (br. s., 1H), 5.20 (d, J=4.5 Hz, 1H), 4.72 (d,J=1.8 Hz, 1H), 4.60 (s, 1H), 4.56 (s, 2H), 4.24-4.15 (m, 2H), 2.99-2.89(m, 1H), 2.88 (s, 3H), 2.83-2.61 (m, 3H), 2.55 (td, J=10.8, 5.5 Hz, 1H),2.31-1.02 (m, 37H), 1.69 (s, 3H), 1.27 (q, J=7.2 Hz, 3H), 1.06 (s, 3H),0.98 (s, 6H), 0.92 (s, 3H), 0.87 (s, 3H); LC/MS m/e 898.7 [(M+H)⁺, calcdfor C₅₄H₇₉N₃O₆S 898.6], t_(R)=4.44 min (LCMS Method 14).

Step 2. To a solution of (S)-ethyl1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1r,4S)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylate(131 mg, 0.146 mmol) in dioxane (4 mL) and MeOH (2 mL) was added lithiumhydroxide (2 mL, 2.00 mmol, 1 M aq). The mixture was heated at 60° C.for 10.5 h. Only a small amount of starting material was detected byLC/MS (LCMS Method 16). The reaction was stopped at this point. Themixture was cooled to room temperature and was partially neutralized bythe addition of 6 N HCl (250 μL). The mixture was then filtered througha syringe filter, and was purified by reverse phase HPLC (5 injections)(Preparative HPLC Method 4). The organic solvent was evaporated on therotovapor and the aqueous mixture was lyophilized to afford(S)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1r,4S)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid, TFA (69 mg, 48% yield) as a white amorphous solid: ¹H NMR (400MHz, Acetic Acid-d₄) δ 8.43 (dd, J=5.0, 2.0 Hz, 1H), 8.06 (dd, J=7.5,2.0 Hz, 1H), 7.12 (dd, J=7.5, 5.3 Hz, 1H), 5.44 (br. s., 1H), 5.27 (d,J=4.8 Hz, 1H), 4.83 (s, 1H), 4.73 (s, 1H), 4.69-4.61 (m, 2H), 3.43-3.29(m, 2H), 3.20-3.10 (m, 1H), 2.99 (s, 3H), 2.91-2.81 (m, J=9.0 Hz, 1H),2.74 (d, J=17.6 Hz, 1H), 2.40-1.33 (m, 37H), 1.76 (s, 3H), 1.15 (s, 3H),1.10 (s, 3H), 1.04 (s, 3H), 1.00 (s, 3H), 0.95 (s, 3H); LC/MS (ESI) m/e870.7 [(M+H) calcd for C₅₂H₇₅N₃O₆S 870.5], t_(R)=2.37 min (LCMS Method15); HPLC (Analytical HPLC Method 3): t_(R)=16.00 min; HPLC (AnalyticalHPLC Method 4): 7R=13.90 min.

Example A5. Preparation of(R)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((ls,4R)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid

Step 1. Preparation of((R)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(ethoxycarbonyl)cyclohex-3-en-1-yl)methylbenzoate

To a flask containing(1R,3aS,5aR,5bR,7aR,11aR,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yltrifluoromethanesulfonate (2.2 g, 3.94 mmol) was added(S)-(1-(ethoxycarbonyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-en-1-yl)methylbenzoate (2.94 g, 7.10 mmol), potassium phosphate tribasic (3.35 g,15.78 mmol), 2-dicyclohexylphosphino-2′,6′-dimethoxybiphenyl (S-Phos)(0.121 g, 0.296 mmol) and palladium(II) acetate (0.044 g, 0.197 mmol).The mixture was diluted with 1,4-dioxane (60 mL) and water (15 mL) andwas flushed with N₂ and heated at 75° C. for 16 h. The mixture wascooled to rt. The mixture was diluted with water (150 mL) and extractedwith ethyl acetate (3×100 mL). The organic layers were washed with brine(200 mL), dried over magnesium sulfate, filtered, and concentrated underreduced pressure. The residue was purified by column chromatography onsilica gel The residue was purified by column chromatography on silicagel (50% ethyl acetate with 4% MeOH and 0.8% aminonium hydroxide/50%hexanes→70% ethyl acetate with 4% MeOH and 0.8% aminonium hydroxide/30%hexanes, 220 g column) to afford((R)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(ethoxycarbonyl)cyclohex-3-en-1-yl)methylbenzoate (2.47 g, 90% yield) as an off-white solid: ¹H NMR (400 MHz,CHLOROFORM-d) δ 8.05-8.00 (m, 2H), 7.61-7.55 (m, 1H), 7.45 (t, J=1.1 Hz,2H), 5.38 (br. s., 1H), 5.25-5.19 (m, 1H), 4.75 (s, 1H), 4.62 (s, 1H),4.46 (q, J=10.8 Hz, 2H), 4.19 (q, J=7.0 Hz, 2H), 2.74-2.66 (m, 1H), 2.56(td, J=10.9, 5.1 Hz, 1H), 2.29-1.96 (m, 6H), 1.87 (dt, J=12.9, 6.2 Hz,1H), 1.78-1.11 (m, 20H), 1.71 (s, 3H), 1.24 (t, J=7.3 Hz, 3H), 1.09 (s,3H), 0.98 (br. s., 3H), 0.97 (br. s., 3H), 0.95 (s, 3H), 0.89 (s, 3H);LC/MS m/e 696.7 [(M+H)⁺, calcd for C₄₆H₆₅NO₄ 696.5], t_(R)=2.55 min(LCMS Method 15).

Step 2. Preparation of (R)-ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(hydroxymethyl)cyclohex-3-enecarboxylate

To a solution of((R)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(ethoxycarbonyl)cyclohex-3-en-1-yl)methylbenzoate (1.20 g, 1.724 mmol in THF (10 mL) and MeOH (1 mL) was addedsodium hydroxide (1.897 mL, 1.897 mmol). The reaction mixture wasstirred at r.t. for 14 h. The solid was removed by filtration. Themixture was transferred to a separatory funnel containing saturatedaqueous NaHCO₃ solution (10 mL)/water (10 mL). The aqueous layer wasextracted with 5% methanol in ethyl acetate (5×25 mL). The combinedorganic layers were washed with brine (10 mL). The brine wash wasreextracted with 5% methanol in ethyl acetate. The combined organiclayers were dried over MgSO₄, filtered, and concentrated to afford(R)-ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(hydroxymethyl)cyclohex-3-enecarboxylate(450 mg, 44% yield) as a white solid. The crude product was useddirectly in the next step. ¹H NMR (400 MHz, CHLOROFORM-d) δ 5.34 (br.s., 1H), 5.20 (dd, J=6.0, 1.8 Hz, 1H), 4.75 (d, J=2.0 Hz, 1H), 4.62 (s,1H), 4.20 (q, J=7.2 Hz, 2H), 3.70 (s, 2H), 2.62-2.51 (m, 2H), 2.21-2.14(m, 2H), 2.10-1.94 (m, 4H), 1.82-1.12 (m, 21H), 1.71 (s, 3H), 1.29 (t,1=1.2 Hz, 3H), 1.09 (s, 3H), 0.98 (s, 3H), 0.97 (s, 3H), 0.95 (s, 3H),0.89 (s, 3H); LC/MS (ESI) m/e 614.6 [(M+H)⁺, calcd for C₃₉H₆₁NO₃Na614.5], t_(R)=4.27 min (LCMS Method 14).

Step 3. Preparation of (R)-ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((3-cyanopyridin-2-yl)oxy)methyl)cyclohex-3-enecarboxylate

To a solution of (R)-ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(hydroxymethyl)cyclohex-3-enecarboxylate(412 mg, 0.696 mmol) and 2-fluoronicotinonitrile (170 mg, 1.392 mmol) inTHF (7 mL) and DMF (1 mL) at 0° C. was added potassium tert-butoxide(0.835 mL, 0.835 mmol). The cooling bath was removed and the reactionmixture was stirred at 20° C. for 1.5 h. The mixture was transferred toa separatory funnel containing saturated aqueous NaHCO₃ solution (15mL). The aqueous layer was extracted with ethyl acetate (4×25 mL). Thecombined organic layers were washed with brine (15 mL), dried overMgSO₄, filtered, and concentrated. The product was purified by columnchromatography on silica gel (50% of a 5% methanol in ethyl acetatesolution/50% hexanes→100% of a 5% methanol in ethyl acetate solution; 40g column) to afford (R)-ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((3-cyanopyridin-2-yl)oxy)methyl)cyclohex-3-enecarboxylate(365 mg, 0.526 mmol, 76% yield) as an off-white solid: ¹H NMR (400 MHz,CHLOROFORM-d) δ 8.34 (dd, J=5.0, 2.0 Hz, 1H), 7.88 (dd, J=7.4, 1.9 Hz,1H), 6.99 (dd, J=7.5, 5.0 Hz, 1H), 5.38 (br. s., 1H), 5.21 (dd, J=6.3,1.8 Hz, 1H), 4.74 (d, J=2.0 Hz, 1H), 4.62 (dd, J=2.1, 1.4 Hz, 1H),4.60-4.52 (m, 2H), 4.19 (qd, J=7.1, 2.5 Hz, 2H), 2.73 (d, J=17.1 Hz,1H), 2.56 (td, J=10.9, 5.4 Hz, 1H), 1.78-1.13 (m, 21H), 2.27-1.87 (m,6H), 1.71 (s, 3H), 1.26 (t, J=6.8 Hz, 3H), 1.09 (s, 3H), 0.98 (s, 3H),0.97 (s, 3H), 0.94 (s, 3H), 0.88 (s, 3H); LC/MS (ESI) m/e 694.7 [(M+H)⁺,calcd for C₄₅H₆₄N₃O₃ 694.5], t_(R)=4.51 min (LCMS Method 14).

Step 4. Preparation of (R)-ethyl1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1s,4R)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylate

(R)-Ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((3-cyanopyridin-2-yl)oxy)methyl)cyclohex-3-enecarboxylate(150 mg, 0.216 mmol) and2-((1s,4s)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)acetaldehyde (76 mg,0.346 mmol) were dissolved in MeOH (1.4 mL) and acetic acid (0.28 mL).Borane-2-picoline complex (37.0 mg, 0.346 mmol) was added and themixture was stirred at room temperature for 14 h. The mixture wastransferred to a separatory funnel containing saturated aqueous sodiumbicarbonate solution (3 mL) and sodium carbonate solution (2 mL). Theaqueous layer was extracted with ethyl acetate (5×10 mL). The combinedorganic layers were washed with brine (5 mL), dried over MgSO₄,filtered, and concentrated. The product was purified by columnchromatography on silica gel (30% ethyl acetate with 5% methanol/70%hexanes→100% ethyl acetate with 5% methanol; 24 g column, 25 mingradient) to afford (R)-ethyl1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1s,4R)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylate(130 mg, 67% yield) as a white foam: ¹H NMR (500 MHz, CHLOROFORM-d) δ8.34 (dd, J=5.0, 2.0 Hz, 1H), 7.88 (dd, J=7.5, 2.0 Hz, 1H), 6.99 (dd,J=7.5, 5.0 Hz, 1H), 5.38 (br. s., 1H), 5.23-5.19 (m, 1H), 4.75 (d, J=1.7Hz, 1H), 4.62 (s, 1H), 4.59-4.52 (m, 2H), 4.19 (dtt, J=10.8, 7.2, 3.8Hz, 2H), 2.85 (s, 3H), 2.83-2.70 (m, 4H), 2.55 (td, J=10.9, 5.6 Hz, 1H),2.28-0.89 (m, 37H), 1.70 (s, 3H), 1.26 (t, J=7.1 Hz, 3H), 1.06 (s, 3H),0.98 (s, 3H), 0.96 (s, 3H), 0.95 (s, 3H), 0.87 (s, 3H); LC/MS (ESI) m/e898.7 [(M+H)⁺, calcd for C₅₄H₈₀N₃O₆S 898.6], t_(R)=4.43 min (LCMS Method14).

Step 5. To a solution of (R)-ethyl1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1s,4R)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylate(124 mg, 0.138 mmol) in dioxane (4 mL) and MeOH (2 mL) was added lithiumhydroxide (2 mL, 2.00 mmol, 1 M aq). The mixture was heated at 60° C.for 10 h. Some starting material starting was detected by LC/MS (LCMSMethod 16) along with formation of an amide by-product due to hydrolysisof the nitrile. The reaction was stopped at this point. The mixture wascooled to room temperature and was partially neutralized by the additionof 6 N HCl (250 μL). The mixture was then filtered through a syringefilter, and was purified by reverse phase HPLC (5 injections)(Preparative HPLC Method 4). The organic solvent was evaporated on therotovapor and the aqueous mixture was lyophilized to afford(R)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1s,4R)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid, TFA (48.1 mg, 34% yield) as a white amorphous solid: NMR (500 MHz,Acetic Acid-d₄) δ 8.42 (dd, J=5.1, 1.9 Hz, 1H), 8.05 (dd, J=7.6, 1.9 Hz,1H), 7.11 (dd, J=7.5, 5.2 Hz, 1H), 5.43 (br. s., 1H), 5.27 (d, J=4.6 Hz,1H), 4.83 (s, 1H), 4.72 (s, 1H), 4.68-4.59 (m, 2H), 3.46-3.33 (m, 2H),3.09-2.99 (m, 1H), 2.96 (s, 3H), 2.89-2.81 (m, 1H), 2.74 (d, J=16.5 Hz,1H), 2.34-1.13 (m, 37H), 1.75 (s, 3H), 1.17 (s, 3H), 1.09 (s, 3H), 1.02(s, 3H), 1.00 (s, 3H), 0.95 (s, 3H); LC/MS (ESI) m/e 870.7 [(M+H)⁺,calcd for C₅₂H₇₆N3O6S 870.5], t_(R)=1.24 min (LCMS Method 16); HPLC(Analytical HPLC Method 3): (R=12.24 min; HPLC (Analytical HPLC Method4): (R=11.77 min.

Example A6. Preparation of(R)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1r,4S)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid

Step 1. Preparation of (R)-ethyl1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1r,4S)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylate

(R)-Ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((3-cyanopyridin-2-yl)oxy)methyl)cyclohex-3-enecarboxylate(150 mg, 0.216 mmol) and2-((1r,4r)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)acetaldehyde (76 mg,0.346 mmol) were dissolved in MeOH (1.6 mL) and acetic acid (0.32 mL).Borane-2-picoline complex (37.0 mg, 0.346 mmol) was added and themixture was stirred at room temperature for 14 h. The mixture wastransferred to a separatory funnel containing saturated aqueous sodiumbicarbonate solution (20 mL). The aqueous layer was extracted with ethylacetate (5×20 mL). The combined organic layers were washed with brine(50 mL), dried over MgSO₄, filtered, and concentrated. The product waspurified by column chromatography on silica gel (30% ethyl acetate with5% methanol/70% hexanes→100% ethyl acetate with 5% methanol; 24 gcolumn) to afford (R)-ethyl1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1r,4S)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylate(131 mg, 68% yield) as a white foam: ¹H NMR (400 MHz, CHLOROFORM-d) δ8.33 (dd, J=5.1, 1.9 Hz, 1H), 7.87 (dd, J=7.4, 1.9 Hz, 1H), 6.98 (dd,J=7.4, 5.1 Hz, 1H), 5.37 (br. s., 1H), 5.22-5.17 (m, 1H), 4.71 (d, J=1.8Hz, 1H), 4.61-4.51 (m, 3H), 4.23-4.14 (m, 2H), 2.99-2.90 (m, 1H), 2.87(s, 3H), 2.82-2.61 (m, 3H), 2.54 (td, J=10.8, 5.5 Hz, 1H), 2.23-1.02 (m,37H), 1.68 (s, 3H), 1.26 (q, J=7.3 Hz, 3H), 1.06 (s, 3H), 0.97 (s, 3H),0.95 (s, 3H), 0.93 (s, 3H), 0.86 (s, 3H); LC/MS m/e 898.7 [(M+H)⁺, calcdfor C₅₄H₇₉N₃O₆S 898.6], t_(R)=4.43 min (LCMS Method 14).

Step 2. To a solution of (R)-ethyl1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1r,4S)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylate(107 mg, 0.119 mmol) in dioxane (4 mL) and MeOH (2 mL) was added lithiumhydroxide (2 mL, 2.00 mmol, 1 M aq). The mixture was heated at 60° C.for 10.5 h. Only a small amount of starting material was detected byLC/MS (LCMS Method 16). The reaction was stopped at this point. Themixture was cooled to room temperature and was partially neutralized bythe addition of 6 N HCl (250 μL). The mixture was then filtered througha syringe filter, and was purified by reverse phase HPLC (5 injections)(Preparative HPLC Method 4). The organic solvent was evaporated on therotovapor and the aqueous mixture was lyophilized to afford(R)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-((1r,4S)-1-hydroxy-4-(methylsulfonyl)cyclohexyl)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid, TFA (58 mg, 49% yield) as a white amorphous solid: ¹H NMR (400MHz, Acetic Acid-d₄) δ 8.43 (dd, J=5.1, 1.9 Hz, 1H), 8.06 (dd, J=7.5,1.8 Hz, 1H), 7.12 (dd, J=7.5, 5.0 Hz, 1H), 5.44 (br. s., 1H), 5.27 (d,J=4.8 Hz, 1H), 4.83 (s, 1H), 4.73 (s, 1H), 4.69-4.60 (m, 2H), 3.43-3.29(m, 2H), 3.20-3.09 (m, 1H), 2.99 (s, 3H), 2.91-2.81 (m, 1H), 2.75 (d,J=15.3 Hz, 1H), 2.32-1.33 (m, 37H), 1.76 (s, 3H), 1.15 (s, 3H), 1.10 (s,3H), 1.03 (s, 3H), 1.00 (s, 3H), 0.95 (s, 3H); LC/MS (ESI) m/e 870.6[(M+H)⁺, calcd for C₅₂H₇₅N3O6S 870.5], t_(R)=2.30 min (LCMS Method 15);HPLC (Analytical HPLC Method 3): t_(R)=14.96 min; HPLC (Analytical HPLCMethod 4): t_(R)=14.64 min.

Example A7 and Example A8. Preparation of(R)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((R)-2,3-dihydroxypropyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid, TFA (Example A7) and(R)-1-(((3-carbamoylpyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((R)-2,3-dihydroxypropyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid (Example A8)

Step 1. Preparation of((R)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(ethoxycarbonyl)cyclohex-3-en-1-yl)methylbenzoate

In a 150 mL medium pressure flask was combined (1R,3aS,5aR,5bR,7aR,11aR,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yltrifluoromethanesulfonate (1.5 g, 2.69 mmol), (R)-(1-(ethoxycarbonyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-en-1-yl)methylbenzoate (1.259 g, 3.04 mmol) and Buchwald pre-catalyst (0.127 g, 0.161mmol) in THF (25 mL). To the reaction mixture was added a solution ofaqueous 0.5 M K₃PO₄ (13.45 mL, 6.72 mmol). The resulting brown solutionwas sparged with N₂(g), stirred at 72° C. overnight. After 16 h, thereaction was allowed to cool to rt, diluted with EtOAc (50 mL) andwashed with 1.5M K₃PO₄ (50 mL). The aqueous layer was extracted with2×50 mL EtOAc. The combined organic layer was washed with brine, driedover MgSO₄, filtered and concentrated to grey foam. Crude material wasdissolved in DCM and loaded onto a silica gel column (SiO₂, 80 g Iscocartridge, eluted with 0% B to 50% B over 4 column volumes, and hold at50% B until all product eluted, solvent A=DCM, solvent B=90:10 DCM:MeOH)and dried in vacuo to give((R)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(ethoxycarbonyl)cyclohex-3-en-1-yl)methylbenzoate (1.8 g, 2.59 mmol, 96% yield) as brown solid. LCMS: m/z 696.6(M+H⁺), retention time 1.589 min (LCMS Method 16). ¹H NMR (400 MHz,CHLOROFORM-d) δ 8.06-7.96 (m, 2H), 7.63-7.53 (m, 1H), 7.48-7.39 (m, 2H),5.36 (br. s., 1H), 5.20 (dd, J=6.1, 1.7 Hz, 1H), 4.73 (d, J=2.0 Hz, 1H),4.61 (s, 1H), 4.44 (q, J=10.8 Hz, 2H), 4.18 (qd, J=7.1, 1.0 Hz, 2H),2.77-2.64 (m, 1H), 2.55 (td, J=10.9, 5.3 Hz, 1H), 2.26-2.13 (m, 3H),2.08 (td, J=12.7, 5.7 Hz, 2H), 2.00 (dd, J=17.0, 6.5 Hz, 1H), 1.85 (dt,J=13.1, 6.4 Hz, 1H), 1.78-1.71 (m, 2H), 1.70 (s, 3H), 1.67-1.56 (m, 6H),1.55-1.49 (m, 4H), 1.48-1.38 (m, 6H), 1.37-1.26 (m, 3H), 1.24-1.19 (m,3H), 1.08 (s, 3H), 0.97 (s, 3H), 0.96 (br. s., 3H), 0.94 (s, 3H), 0.87(s, 3H).

Step 2. Preparation of (R)-ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(hydroxymethyl)cyclohex-3-enecarboxylate

To a solution of((R)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(ethoxycarbonyl)cyclohex-3-en-1-yl)methylbenzoate (0.692 g, 0.994 mmol) in THF (10 mL) and MeOH (1 mL) was addedsodium hydroxide (0.994 mL, 0.994 mmol) and the resulting mixture wasstirred at rt. After 3 h, the reaction was concentrated to dryness andthe material was dissolved in DCM:MeOH and purified by flash columnchromatography (SiO₂, 40 g Isco cartridge, eluted with 95:5 DCM:MeOH)and dried in vacuo to give (R)-ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(hydroxymethyl)cyclohex-3-enecarboxylate(427 mg, 0.721 mmol, 72.6% yield) as light yellow solid. LCMS: m/z 592.5(M+H⁺), retention time 1.705 min (LCMS Method 16). ¹H NMR (400 MHz, 1:1CDCl3:METHANOL-d₄) δ 5.30 (br. s., 1H), 5.14 (d, J=4.6 Hz, 1H), 4.72(br. s., 1H), 4.60 (br. s., 1H), 4.22-4.00 (m, 2H), 3.74-3.53 (m, 2H),2.60-2.42 (m, 2H), 2.13 (br. s., 2H), 2.06-1.87 (m, 4H), 1.78-1.70 (m,1H), 1.67 (br. s., 5H), 1.63-1.51 (m, 6H), 1.43 (br. s., 7H), 1.32 (br.s., 1H), 1.24 (t, J=7.0 Hz, 4H), 1.06 (br. s., 4H), 0.97 (br. s., 3H),0.92 (br. s., 3H), 0.90 (br. s., 3H), 0.85 (br. s., 3H).

Step 3. Preparation of (R)-ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((3-cyanopyridin-2-yl)oxy)methyl)cyclohex-3-enecarboxylate

(R)-ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(hydroxymethyl)cyclohex-3-enecarboxylate(420 mg, 0.710 mmol) and 3-cyano-2-fluoropyridine (130 mg, 1.064 mmol)were combined in DMF (3 mL) and THF (3 mL) chilled to 0° C. To theyellow slurry was treated with a solution of potassium tert-butoxide(0.781 mL, 0.781 mmol) in THF. The reaction became almost totallyhomogeneous; the cold bath was removed and the reaction was stirred tort. After 3.5 h, there was still a small amount of starting materialleft; thus to the reaction was added more 3-cyano-2-fluoropyridine (43.3mg, 0.355 mmol) and potassium tert-butoxide (0.142 mL, 0.142 mmol) andstirred at RT for an additional 1 h. The reaction was diluted with EtOAcand washed with 0.5N HCl 25 mL. The aqueous layer was extracted with2×50 mL EtOAc. The combined organic layer was washed with saturatedNaHCO₃, brine, dried over MgSO₄, filtered and concentrated to brownpaste. Crude material was purified by flash column chromatography (SiO₂,40 g Isco cartridge, eluted with 95:5 DCM:MeOH) and dried under vacuo togive (R)-ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((3-cyanopyridin-2-yl)oxy)methyl)cyclohex-3-enecarboxylate(426 mg, 0.614 mmol, 87% yield) as light brown solid. LCMS: m/z 694.9(M+H⁺), retention time 1.517 min (LCMS Method 16).

Step 4. Preparation of (R)-ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((R)-3-((tert-butyldimethylsilyl)oxy)-2-hydroxypropyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((3-cyanopyridin-2-yl)oxy)methyl)cyclohex-3-enecarboxylate,TFA

To a solution of (R)-ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((3-cyanopyridin-2-yl)oxy)methyl)cyclohex-3-enecarboxylate(48.5 mg, 0.070 mmol) in acetonitrile (0.5 mL) and 1,4-dioxane (0.5 mL)was added tert-butyldimethylsilyl (R)-(−)-glycidyl ether (0.094 mL,0.489 mmol) and the mixture was stirred at 100° C. overnight. After 19h, the reaction was allowed to cool to RT and was purified by reversephase preparative HPLC using preparative HPLC method 8 and dried undervacuo to give (R)-ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((R)-3-((tert-butyldimethylsilyl)oxy)-2-hydroxypropyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((3-cyanopyridin-2-yl)oxy)methyl)cyclohex-3-enecarboxylate,TFA (22.8 mg, 0.023 mmol, 32.7% yield, 53.5% yield based on recoveredstarting material) and recovered starting material (21.9 mg), both asclear glass solid. LCMS: m/z 882.4 (M+H⁺), retention time 1.849 min(LCMS Method 16).

Step 5. To a solution of (R)-ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((R)-3-((tert-butyldimethylsilyl)oxy)-2-hydroxypropyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((3-cyanopyridin-2-yl)oxy)methyl)cyclohex-3-enecarboxylate,TFA (22.8 mg, 0.023 mmol) in 2-Me-THF (1 mL) and H₂O (0.3 mL) was addeda solution of tetrabutylaminonium hydroxide (0.105 mL, 0.160 mmol) andthe mixture was stirred at RT for 4 h but LC/MS showed no reaction. Thereaction was then stirred at 50° C. After 14 h, LC/MS showedapproximately 60% of starting material remained; thus the mixture wasstirred at 50° C. for another night. After 40 h, the reaction mixturewas purified by reverse phase preparative HPLC using preparative HPLCmethod 8 and product fractions were dried in vacuo to give two products,both as glass solids. Example A8 was the first of the two isolatedproducts to elute from the preparative HPLC column:(R)-1-(((3-carbamoylpyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((R)-2,3-dihydroxypropyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid, TFA (4.0 mg, 4.36 μmol, 19.04% yield). LCMS: m/z 758.7 (M+H⁺),retention time 1.219 min (LCMS Method 16). ¹H NMR (400 MHz, 1:1CDCl₃:METHANOL-d₄) δ 8.47-8.36 (m, 1H), 8.25 (d, J=3.2 Hz, 1H), 7.08(dd, J=7.6, 4.9 Hz, 1H), 5.34 (br. s., 1H), 5.19 (d, J=4.9 Hz, 1H), 4.79(s, 1H), 4.71 (br. s., 1H), 4.06-3.90 (m, 1H), 3.66 (d, J=4.2 Hz, 2H),3.23-3.11 (m, 1H), 3.03-2.92 (m, 1H), 2.80-2.61 (m, 2H), 2.48-1.90 (m,10H), 1.84 (d, J=6.6 Hz, 1H), 1.71 (s, 4H), 1.69-1.21 (m, 15H), 1.15 (d,J=12.7 Hz, 1H), 1.08 (s, 3H), 1.05 (s, 3H), 0.96 (s, 3H), 0.92 (s, 3H),0.86 (s, 3H).

Example A7 was the second of the two isolated products to elute from thepreparative HPLC column:(R)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((R)-2,3-dihydroxypropyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid, TFA (6.5 mg, 7.46 μmol, 32.6% yield). LCMS: m/z 740.6 (M+H⁺),retention time 1.289 min (LCMS Method 16). ¹H NMR (400 MHz, 1:1CDCl₃:METHANOL-d₄) δ 8.33 (dd, J=5.0, 1.8 Hz, 1H), 7.94 (dd, J=7.6, 1.7Hz, 1H), 7.03 (dd, J=7.6, 5.1 Hz, 1H), 5.34 (br. s., 1H), 5.18 (d, J=4.9Hz, 1H), 4.79 (s, 1H), 4.71 (s, 1H), 3.99 (dd, J=8.6, 3.9 Hz, 1H), 3.66(d, J=4.2 Hz, 2H), 3.18 (dd, J=2.1, 3.5 Hz, 1H), 2.98 (dd, J=11.9, 8.9Hz, 1H), 2.78-2.56 (m, 2H), 2.35-2.08 (m, 4H), 2.08-1.87 (m, 6H), 1.75(br. s., 1H), 1.72 (s, 3H), 1.70-1.53 (m, 6H), 1.51-1.22 (m, 8H),1.21-1.12 (m, 1H), 1.08 (s, 3H), 1.05 (s, 3H), 0.96 (s, 3H), 0.91 (s,3H), 0.86 (s, 3H).

Example A9 and Example A10. Preparation of(R)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((R)-2-hydroxy-3-methoxypropyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid (Example A9) and(R)-1-(((3-carbamoylpyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((R)-2-hydroxy-3-methoxypropyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid (Example A10)

The title compounds were prepared in 7.1% and 16.1% yield, respectively,from (R)-ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((3-cyanopyridin-2-yl)oxy)methyl)cyclohex-3-enecarboxylatefollowing the same procedure as described for the preparation of(R)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((R)-2,3-dihydroxypropyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid, TFA and(R)-1-(((3-carbamoylpyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((R)-2,3-dihydroxypropyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid, except (R)-(−)-methyl glycidyl ether was used instead oftert-butyldimethylsilyl (R)-(−)-glycidyl ether in Step 4. Example A10was the first of the two isolated products to elute from the preparativeHPLC column:(R)-1-(((3-carbamoylpyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((R)-2-hydroxy-3-methoxypropyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid (5.6 mg, 7.25 μmol, 16.13% yield). LCMS: m/e 772.6 (M+H⁺), 1.284min (LCMS Method 16). Example A9 was the second of the two isolatedproducts to elute from the preparative HPLC column:(R)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((R)-2-hydroxy-3-methoxypropyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid, TFA (2.9 mg, 3.17 μmol, 7.06% yield). LCMS: m/z 754.6 (M+H⁺),retention time 1.345 min (LCMS Method 16). ¹H NMR (400 MHz, 1:1CDCl₃:METHANOL-d₄) δ 8.33 (dd, J=5.0, 1.8 Hz, 1H), 7.94 (dd, J=7.5, 1.8Hz, 1H), 7.03 (dd, J=7.5, 5.0 Hz, 1H), 5.34 (br. s., 1H), 5.18 (d, J=4.4Hz, 1H), 4.79 (s, 1H), 4.71 (s, 1H), 4.07 (dd, J=9.9, 4.0 Hz, 1H),3.54-3.44 (m, 2H), 3.39 (s, 3H), 3.15 (dd, J=11.9, 3.3 Hz, 1H), 2.93 (t,J=11.1 Hz, 1H), 2.75-2.59 (m, 2H), 2.31-2.08 (m, 4H), 2.07-1.89 (m, 6H),1.79-1.73 (m, 1H), 1.71 (s, 3H), 1.67 (br. s., 1H), 1.65-1.57 (m, 3H),1.56-1.39 (m, 6H), 1.37-1.22 (m, 4H), 1.21-1.13 (m, 1H), 1.08 (s, 3H),1.05 (s, 3H), 0.96 (s, 3H), 0.91 (s, 3H), 0.86 (s, 3H).

Example A11 and Example A12. Preparation of(R)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((S)-2-hydroxy-3-methoxypropyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid, TFA (Example A11) and(R)-1-(((3-carbamoylpyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((S)-2-hydroxy-3-methoxypropyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid, TFA (Example A12)

The title compounds were prepared in 26.9% and 6.1% yield, respectively,from (R)-ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((3-cyanopyridin-2-yl)oxy)methyl)cyclohex-3-enecarboxylatefollowing the same procedure as described for the preparation of(R)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((R)-2,3-dihydroxypropyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid, TFA and(R)-1-(((3-carbamoylpyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((R)-2,3-dihydroxypropyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid, except (S)-(+)-methyl glycidyl ether was used instead oftert-butyldimethylsilyl (R)-(−)-glycidyl ether in Step 4. Example A12was the first of the two isolated products to elute from the preparativeHPLC column:(R)-1-(((3-carbamoylpyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((S)-2-hydroxy-3-methoxypropyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid, TFA (2.2 mg, 2.359 μmol, 6.11% yield). LCMS: m/z 772.6 (M+H⁺),retention time 1.279 min (LCMS Method 16). ¹H NMR (400 MHz, 1:1CDCl3:METHANOL-d4) δ 8.41 (dd, J=7.6, 2.0 Hz, 1H), 8.25 (dd, J=4.8, 2.1Hz, 1H), 7.08 (dd, J=7.6, 4.9 Hz, 1H), 5.35 (br. s., 1H), 5.19 (d, J=4.6Hz, 1H), 4.80 (s, 1H), 4.73 (s, 1H), 4.11 (t, J=4.0 Hz, 1H), 3.69-3.63(m, 1H), 3.61-3.55 (m, 1H), 3.44 (s, 3H), 3.27-3.20 (m, 1H), 3.19-3.12(m, 1H), 2.72 (d, J=15.9 Hz, 1H), 2.63-2.52 (m, 1H), 2.26 (br. s., 1H),2.22-2.08 (m, 4H), 2.07-1.95 (m, 4H), 1.88-1.74 (m, 3H), 1.72 (s, 3H),1.70-1.62 (m, 2H), 1.62-1.41 (m, 8H), 1.41-1.22 (m, 4H), 1.16 (br. s.,1H), 1.09 (s, 3H), 1.05 (s, 3H), 0.96 (s, 3H), 0.92 (s, 3H), 0.87 (s,3H). Example A11 was the second of the two isolated products to elutefrom the preparative HPLC column:(R)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((S)-2-hydroxy-3-methoxypropyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid, TFA (9.5 mg, 10.40 μmol, 26.9% yield). LCMS: m/z 754.6 (M+H⁺),retention time 1.347 min (LCMS Method 16). ¹H NMR (400 MHz, 1:1CDCl₃:METHANOL-d₄) δ 8.33 (dd, J=5.1, 2.0 Hz, 1H), 7.94 (dd, J=7.6, 2.0Hz, 1H), 7.03 (dd, J=7.6, 4.9 Hz, 1H), 5.34 (br. s., 1H), 5.18 (d, J=4.4Hz, 1H), 4.80 (s, 1H), 4.73 (s, 1H), 4.11 (t, J=4.0 Hz, 1H), 3.70-3.54(m, 2H), 3.44 (s, 3H), 3.27-3.20 (m, 1H), 3.19-3.12 (m, 1H), 2.64 (d,J=15.9 Hz, 1H), 2.60-2.51 (m, 1H), 2.20 (d, J=16.6 Hz, 3H), 2.11-1.89(m, 7H), 1.82-1.74 (m, 2H), 1.72 (s, 3H), 1.70-1.63 (m, 2H), 1.63-1.22(m, 12H), 1.20-1.11 (m, 1H), 1.09 (s, 3H), 1.05 (s, 2H), 0.96 (s, 3H),0.92 (s, 3H), 0.87 (s, 3H). ¹³C NMR (101 MHz, 1:1 CDCl3:METHANOL-d₄) δ178.3, 164.5, 152.27-152.01, 148.9, 147.7, 144.1, 139.7, 122.7, 121.9,117.7, 112.5, 97.5, 78.5, 76.6, 72.6, 71.5, 65.1, 60.2, 53.8, 50.0,46.8, 46.6, 45.3, 42.8, 41.4, 38.6, 38.3, 36.9, 34.2, 32.6, 31.1, 30.2,30.1, 28.1, 27.4, 26.8, 25.9, 22.0, 21.7, 20.3, 19.2, 17.0, 16.0, 15.0.

Example A13 and Example A14 Preparation of(R)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((R)-2,3-dihydroxy-2-methylpropyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid, TFA (Example A13) and(R)-1-(((3-carbamoylpyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((R)-2,3-dihydroxy-2-methylpropyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid, TFA (Example A14)

The title compounds were prepared in 26.0% and 13.6% yield,respectively, from (R)-ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((3-cyanopyridin-2-yl)oxy)methyl)cyclohex-3-enecarboxylatefollowing the same procedure as described for the preparation of(R)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((R)-2,3-dihydroxypropyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid, TFA and(R)-1-(((3-carbamoylpyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((R)-2,3-dihydroxypropyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid, except (2R)-(−)-2-methylglycidyl 4-nitrobenzoate was used insteadof tert-butyldimethylsilyl (R)-(−)-glycidyl ether in Step 4.

Example A14 was the first of the two isolated products to elute from thepreparative HPLC column:(R)-1-(((3-carbamoylpyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((R)-2,3-dihydroxy-2-methylpropyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid, TFA (4.8 mg, 5.91 μmol, 13.57% yield). LCMS: m/z 772.6 (M+H⁺),retention time 1.242 min (LCMS Method 16). ¹H NMR (400 MHz, 1:1CDCl₃:METHANOL-d₄) δ 8.41 (dd, J=7.6, 2.0 Hz, 1H), 8.25 (dd, J=4.9, 2.0Hz, 1H), 7.08 (dd, J=7.6, 4.9 Hz, 1H), 5.34 (br. s., 1H), 5.19 (d, J=4.6Hz, 1H), 4.81 (s, 1H), 4.73 (s, 1H), 3.69 (s, 2H), 2.97 (d, J=12.2 Hz,1H), 2.79-2.68 (m, 1H), 2.68-2.59 (m, 1H), 2.34-2.23 (m, 1H), 2.22-2.08(m, 3H), 2.08-1.95 (m, 4H), 1.90-1.80 (m, 1H), 1.79-1.74 (m, 1H), 1.73(s, 3H), 1.71-1.66 (m, 1H), 1.66-1.53 (m, 4H), 1.52-1.33 (m, 6H), 1.30(br. s., 2H), 1.23 (s, 3H), 1.20-1.09 (m, 2H), 1.06 (s, 3H), 1.04 (s,3H), 0.96 (s, 3H), 0.92 (s, 3H), 0.86 (s, 3H).

Example A13 was the second of the two isolated products to elute fromthe preparative HPLC column:(R)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((R)-2,3-dihydroxy-2-methylpropyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid, TFA (8.7 mg, 0.011 mmol, 26.0% yield). LCMS: m/z 754.6 (M+H⁺),retention time 1.309 min (LCMS Method 16). ¹H NMR (400 MHz, 1:1CDCl₃:METHANOL-d₄) δ 8.33 (dd, J=5.0, 1.8 Hz, 1H), 7.94 (dd, J=7.6, 2.0Hz, 1H), 7.03 (dd, J=7.6, 5.1 Hz, 1H), 5.34 (br. s., 1H), 5.18 (d, J=4.4Hz, 1H), 4.81 (s, 1H), 4.73 (s, 1H), 3.69 (s, 2H), 2.97 (d, J=12.0 Hz,1H), 2.72-2.59 (m, 2H), 2.30-2.15 (m, 3H), 2.12-1.99 (m, 5H), 1.98-1.88(m, 2H), 1.81-1.74 (m, 1H), 1.73 (s, 3H), 1.71-1.66 (m, 1H), 1.66-1.53(m, 5H), 1.52-1.33 (m, 7H), 1.31-1.25 (m, 1H), 1.23 (s, 3H), 1.19-1.09(m, 2H), 1.06 (s, 3H), 1.04 (s, 2H), 0.96 (s, 3H), 0.91 (s, 3H), 0.86(s, 3H). ¹³C NMR (101 MHZ, 1:1 CDCl3:METHANOL-d₄) δ 178.3, 164.5, 152.2,148.9, 147.8, 144.1, 139.7, 122.7, 121.9, 117.7, 112.4, 97.5, 78.6,72.2, 71.6, 71.5, 69.6, 53.8, 50.0, 46.3, 45.3, 42.9, 42.6, 41.4, 38.6,38.3, 36.9, 34.2, 32.1, 31.1, 30.2, 30.08-30.04, 28.5, 28.1, 27.7, 26.9,26.0, 23.5, 22.0, 21.7, 20.3, 19.4, 17.0, 16.0, 15.0.

Example A15 and Example A16. Preparation of(R)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((R)-2-carboxy-2-hydroxyethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((3-cyanopyridin-2-yl)oxy)methyl)cyclohex-3-enecarboxylicacid, TFA (Example A15) and2-(((R)-1-carboxy-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((R)-2-carboxy-2-hydroxyethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-en-1-yl)methoxy)nicotinicacid, TFA (Example A16)

The title compounds were prepared in 19.5% and 17.9% yield,respectively, from (R)-ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((3-cyanopyridin-2-yl)oxy)methyl)cyclohex-3-enecarboxylatefollowing the same procedure as described for the preparation of(R)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((R)-2,3-dihydroxypropyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid, TFA and(R)-1-(((3-carbamoylpyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((R)-2,3-dihydroxypropyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid, except (R)-methylglycidate was used instead oftert-butyldimethylsilyl (R)-(−)-glycidyl ether in Step 4. Example A16was the first of the two isolated products to elute from the preparativeHPLC column:2-(((R)-1-carboxy-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((R)-2-carboxy-2-hydroxyethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-en-1-yl)methoxy)nicotinicacid (6.0 mg, 7.37 μmol, 17.94% yield). LCMS: m/z 773.5 (M+H⁺),retention time 1.224 min (LCMS Method 16).

¹H NMR (400 MHz, 1:1 CDCl₃:METHANOL-d₄) δ 8.41 (dd, J=7,6, 2.0 Hz, 1H),8.25 (dd, J=4.9, 2.0 Hz, 1H), 7.08 (dd, J=7.6, 4.9 Hz, 1H), 5.35 (br.s., 1H), 5.19 (d, J=4.6 Hz, 1H), 4.80 (s, 1H), 4.71 (s, 1H), 4.44 (dd,J=10.0, 4.2 Hz, 1H), 3.40-3.34 (m, 1H), 3.06 (t, J=1.0 Hz, 1H),2.77-2.62 (m, 2H), 2.26 (br. s., 1H), 2.22-2.05 (m, 5H), 2.04-1.94 (m,3H), 1.89-1.74 (m, 3H), 1.72 (s, 3H), 1.69-1.57 (m, 4H), 1.57-1.40 (m,5H), 1.39-1.22 (m, 4H), 1.22-1.12 (m, 1H), 1.10 (s, 3H), 1.05 (s, 3H),0.96 (s, 3H), 0.92 (s, 3H), 0.87 (s, 3H).

Example A15 was the second of the two isolated products to elute fromthe preparative HPLC column:(R)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((R)-2-carboxy-2-hydroxyethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((3-cyanopyridin-2-yl)oxy)methyl)cyclohex-3-enecarboxylicacid (6.3 mg, 8.02 μmol, 19.52% yield). LCMS: m/z 754.6 (M+H⁺),retention time 1.289 min (LCMS Method 16). ¹H NMR (400 MHz, 1:1CDCl₃:METHANOL-d₄) δ 8.33 (dd, J=5.0, 1.8 Hz, 1H), 7.94 (dd, J=7.6, 2.0Hz, 1H), 7.03 (dd, J=7.5, 5.0 Hz, 1H), 5.34 (br. s., 1H), 5.18 (d, J=4.4Hz, 1H), 4.80 (s, 1H), 4.71 (s, 1H), 4.41 (d, J=5.4 Hz, 1H), 3.06 (t,J=10.8 Hz, 1H), 2.65 (d, J=19.1 Hz, 2H), 2.20 (d, J=15.9 Hz, 3H),2.14-2.05 (m, 2H), 2.05-1.87 (m, 5H), 1.82-1.69 (m, 5H), 1.68-1.57 (m,4H), 1.56-1.41 (m, 5H), 1.39-1.22 (m, 4H), 1.21-1.12 (m, 1H), 1.10 (s,3H), 1.05 (s, 3H), 0.96 (s, 3H), 0.92 (s, 3H), 0.87 (s, 3H).

Example A17 and Example A18. Preparation of(R)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((S)-2-carboxy-2-hydroxyethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((3-cyanopyridin-2-yl)oxy)methyl)cyclohex-3-enecarboxylicacid, TFA (Example A17) and(R)-3-(((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-9-((S)-4-(((3-carbamoylpyridin-2-yl)oxy)methyl)-4-(ethoxycarbonyl)cyclohex-1-en-1-yl)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-3a-yl)amino)-2-hydroxypropanoicacid, TFA (Example A18)

The title compounds were prepared in 19.5% and 16.0% yield,respectively, from (R)-ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((3-cyanopyridin-2-yl)oxy)methyl)cyclohex-3-enecarboxylatefollowing the same procedure as described for the preparation of(R)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((R)-2,3-dihydroxypropyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid, TFA and(R)-1-(((3-carbamoylpyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((R)-2,3-dihydroxypropyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid, except (S)-methylglycidate was used instead oftert-butyldimethylsilyl (R)-(−)-glycidyl ether in Step 4. Example A18was the first of the two isolated products to elute from the preparativeHPLC column:(R)-3-(((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-9-((S)-4-(((3-carbamoylpyridin-2-yl)oxy)methyl)-4-(ethoxycarbonyl)cyclohex-1-en-1-yl)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-3a-yl)amino)-2-hydroxypropanoicacid, TFA (5.7 mg, 6.11 μmol, 15.98% yield). LCMS: m/z 772.7 (M+H⁺),retention time 1.222 min (LCMS Method 16). ¹HNMR (400 MHz, 1:1CDCl₃:METHANOL-d₄) δ 8.41 (dd, J=7.6, 2.0 Hz, 1H), 8.25 (dd, J=4.9, 2.0Hz, 1H), 7.08 (dd, J=7.6, 4.9 Hz, 1H), 5.35 (br. s., 1H), 5.19 (d, J=4.6Hz, 1H), 4.79 (s, 1H), 4.71 (s, 1H), 4.41 (t, J=6.6 Hz, 1H), 3.20 (d,J=6.4 Hz, 2H), 2.71 (d, J=13.4 Hz, 2H), 2.36-2.23 (m, 1H), 2.16 (d,J=14.9 Hz, 2H), 2.13-2.06 (m, 2H), 2.05-1.95 (m, 4H), 1.89-1.80 (m, 1H),1.79-1.74 (m, 1H), 1.72 (s, 3H), 1.68 (br. s., 2H), 1.65-1.57 (m, 2H),1.57-1.50 (m, 2H), 1.49-1.39 (m, 4H), 1.39-1.22 (m, 4H), 1.12 (s, 3H),1.08 (d, J=9.5 Hz, 1H), 1.04 (s, 3H), 0.96 (s, 3H), 0.92 (s, 3H), 0.87(s, 3H).

Example A17 was the second of the two isolated products to elute fromthe preparative HPLC column:(R)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((S)-2-carboxy-2-hydroxyethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((3-cyanopyridin-2-yl)oxy)methyl)cyclohex-3-enecarboxylicacid, TFA (6.8 mg, 7.44 μmol, 19.46% yield). LCMS: m/z 754.6 (M+H⁺),retention time 1.284 min (LCMS Method 16). ¹H NMR (400 MHz, 1:1CDCl₃:METHANOL-d₄) S 8.33 (dd, J=5.0, 1.8 Hz, 1H), 7.94 (dd, J=7.6, 2.0Hz, 1H), 7.03 (dd, J=7.6, 5.1 Hz, 1H), 5.34 (br. s., 1H), 5.18 (d, J=4.6Hz, 1H), 4.79 (br. s., 1H), 4.71 (br. s., 1H), 4.41 (br. s., 1H), 3.20(d, J=5.1 Hz, 2H), 2.70 (br. s., 1H), 2.64 (d, J=18.8 Hz, 1H), 2.20 (d,J=16.1 Hz, 3H), 2.12-1.89 (m, 7H), 1.75 (br. s., 2H), 1.72 (s, 3H),1.70-1.51 (m, 6H), 1.51-1.39 (m, 4H), 1.38-1.22 (m, 4H), 1.12 (s, 3H),1.08 (d, J=9.0 Hz, 1H), 1.04 (s, 3H), 0.96 (s, 3H), 0.92 (s, 3H), 0.87(s, 3H).

Example A19 Preparation of(1R)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-carboxy-2-hydroxypropyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((3-cyanopyridin-2-yl)oxy)methyl)cyclohex-3-enecarboxylicacid

The title compound was prepared in 19.4% yield from (R)-ethyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-amino-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(((3-cyanopyridin-2-yl)oxy)methyl)cyclohex-3-enecarboxylatefollowing the same procedure as described for the preparation of(R)-1-(((3-cyanopyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((R)-2,3-dihydroxypropyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid, TFA and(R)-1-(((3-carbamoylpyridin-2-yl)oxy)methyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-(((R)-2,3-dihydroxypropyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-enecarboxylicacid; except methyl 2-methylglycidate was used instead oftert-butyldimethylsilyl (R)-(−)-glycidyl ether in Step 4. LCMS: m/z768.5 (M+H⁺), retention time 1.295 min (LCMS Method 16). ¹H NMR (400MHz, 1:1 CDCl₃:METHANOL-d₄) δ 8.33 (dd, J=5.0, 1.8 Hz, 1H), 7.94 (dd,J=7.6, 2.0 Hz, 1H), 7.03 (dd, J=7.5, 5.0 Hz, 1H), 5.34 (br. s., 1H),5.18 (d, J=4.6 Hz, 1H), 4.79 (br. s., 1H), 4.71 (br. s., 1H), 3.08-2.89(m, 1H), 2.80-2.57 (m, 2H), 2.33-2.09 (m, 4H), 2.08-1.87 (m, 6H),1.82-1.74 (m, 1H), 1.72 (s, 3H), 1.70-1.57 (m, 4H), 1.56-1.41 (m, 8H),1.40-1.22 (m, 4H), 1.12 (s, 1H), 1.09 (br. s., 1.5H), 1.07 (br. s.,1.5H), 1.05 (s, 3H), 1.01 (s, 1H), 0.96 (s, 3H), 0.92 (s, 3H), 0.86 (br.s., 3H).

Preparation of8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2-oxaspiro[4.5]dec-7-en-1-one

Step 1: Preparation of8-((trimethylsilyl)oxy)-2-oxaspiro[4.5]dec-7-en-1-one

To a 350 mL Chemglass pressure vessel with threaded stopper was added3-methylenedihydrofuran-2(3H)-one (4.31 g, 43.9 mmol) and(buta-1,3-dien-2-yloxy)trimethylsilane (7.50 g, 52.7 mmol) and benzene(100 mL). Hydroquinone (0.726 g, 6.59 mmol) was added, then the solutionwas flushed with nitrogen, sealed and heated to 123° C. for 20 h. Anadditional 2.4 equivalents of (buta-1,3-dien-2-yloxy)trimethylsilane(15.0 g, 105.4 mmol) was then added to the vessel, and the mixture washeated to 123° C. for an additional 60 h. The mixture was concentratedin vacuo to give approximately 19 g of yellow oil. The crude mixture wasloaded with minimum DCM and hexanes onto a hexanes preequilibrated Isco330 g silica cartridge. Elution gradient 100% hexanes to 11.1hexanes:EtOAc over 2 column volumes, then hold 11:1 hex:EtOAc for 3column volumes, then gradient to 5:1 hex:EtAc over 2 column volumes,then hold 5:1 hex:EtOAc for 6 column volumes. Concentration of combinedfractions containing the desired material provided the product as awhite solid: 7.50 g (71.0% yield). ¹H NMR (400 MHz, CHLOROFORM-d) δ 4.85(d, J=5.6 Hz, 1H), 4.40-4.23 (m, 2H), 2.47 (dd, J=16.6, 2.2 Hz, 1H),2.19-2.10 (m, 4H), 2.06 (d, J=3.4 Hz, 1H), 2.04-1.99 (m, 1H), 1.75-1.65(m, 1H), 0.22 (s, 9H).

Step 2. Preparation of 2-oxaspiro[4.5]decane-1,8-dione

8-((trimethylsilyl)oxy)-2-oxaspiro[4.5]dec-7-en-1-one (7.50 g, 31.2mmol) was combined with THF (100 mL) and hydrochloric acid, 0.05Maqueous (3.12 mL, 0.156 mmol). The mixture was stirred for 18 h at RT.The reaction mixture was then concentrated in vacuo to a residue. Theresidue was taken up in EtOAc (200 mL) and washed with saturated NaHCO₃(50 mL) and with brine (50 mL). The organic phase was dried overanhydrous magnesium sulfate, filtered and concentrated in vacuo. Thecrude mixture was loaded in minimum DCM onto a hexanes preequilibratedIsco 330 g silica cartridge. Elution gradient 100% hexanes to 1:1hexanes:EtOAc over 10 column volumes, hold 1:1 hexanes:EtOAc for 6column volumes. Partial separation of the two materials was achieved.Like fractions were combined and set aside, and mixed fractions wererechromatographed in a similar manner. The desired material was themajor product from the reaction and was the second of the two materialsto elute from the silica column. The desired material was recovered as awhite solid: 4.14 g (79.0% yield). ¹H NMR (400 MHz, CHLOROFORM-d) δ 4.40(t, J=7.1 Hz, 2H), 2.87-2.70 (m, 2H), 2.44-2.29 (m, 4H), 2.24 (ddd,J=13.6, 8.3, 5.5 Hz, 2H), 1.96 (dt, J=13.6, 6.5 Hz, 2H).

Step 3. Preparation of 1-oxo-2-oxaspiro[4.5]dec-7-en-8-yltrifluoromethanesulfonate

In a 250 mL round bottom flask fitted with magnetic stirrer and rubberseptum were combined 2-oxaspiro[4.5]decane-1,8-dione (4.13 g, 24.6 mmol)and N,N-bis(trifluoromethylsulfonyl)aniline (10.1 g, 28.2 mmol) inanhydrous tetrahydrofuran (100 mL). The solution was cooled to −78° C.in a dry ice/acetone bath. To the cold solution was added dropwisepotassium hexamethyldisilazide, 0.5M in toluene (56.5 mL, 28.2 mmol)over 15 min. The mixture was stirred at −78° C. for a total of 4 h whenit was treated slowly with 100 mL of saturated aqueous aminoniumchloride. The mixture was stirred at RT for 15 min and was concentratedin vacuo to remove most of the THF, then to the residue was added ethylacetate (300 mL). The resulting mixture was shaken and phases wereseparated. The organic was washed with water (2×100 mL) and with brine(50 mL). The organic was dried over anhydrous magnesium sulfate,filtered and concentrated in vacuo to give a crude yellow oil. The cruderesidue was loaded as an oil onto a hexanes preequilibrated Isco 220 gsilica cartridge and the flask was rinsed with minimum DCM and this wasadded to the column as well. Elution gradient 100% hexanes to 3:1hexanes:EtOAc over 3 column volumes, then hold 3:1 hex:EtOAc for 3column volumes, then 2:1 hex:EtOAc for 3 column volumes. Like productfractions were combined and concentrated in vacuo to give the desiredmaterial as a slightly yellow oil: 6.44 g (87.0% yield). ¹H NMR (400MHz, CHLOROFORM-d) δ 5.86-5.76 (m, 1H), 4.44-4.29 (m, 2H), 2.63 (dd,J=17.7, 2.8 Hz, 1H), 2.59-2.38 (m, 2H), 2.30-2.16 (m, 3H), 2.16-2.04 (m,1H), 1.86 (dt, J=13.7, 2.9 Hz, 1H).

Step 4. In a 250 mL round bottom flask fitted with a reflux condenserwere combined 1-oxo-2-oxaspiro[4.5]dec-7-en-8-yltrifluoromethanesulfonate (6.43 g, 21.4 mmol), potassium acetate (5.25g, 53.5 mmol),4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane) (5.71 g,22.5 mmol) and PdCl₂(dppf).CH₂Cl₂ (0.529 g, 0.642 mmol) in dry1,4-dioxane (100 mL). The mixture was flushed with nitrogen and heatedto 70° C. for 5 h. The reaction mixture was concentrated in vacuo toapprox. 25 mL total volume and was diluted with ethyl acetate (300 mL)and water (150 mL). The mixture was shaken and phases were separated.The organic was again washed with water (100 mL) and then with brine(100 mL). The organic phase was dried over anhydrous magnesium sulfate,filtered and concentrated in vacuo to a deep red residue. The crudemixture was dissolved in minimum DCM and loaded onto a hexanespreequilibrated Isco 220 g silica cartridge. Elution gradient 100%hexanes to 20% ethyl acetate in hexanes over 10 column volumes, thenhold 20% ethyl acetate in hexanes for 6 column volumes, then gradient to15% ethyl acetate in hexanes over 2 column volumes, then hold 25% ethylacetate in hexanes for 6 column volumes. Product fractions were combinedand concentrated in vacuo to give the desired material as a white foamsolid=4.94 g (83.0% yield). ¹H NMR (400 MHz, CHLOROFORM-d) δ 6.60-6.49(m, 1H), 4.39-4.22 (m, 2H), 2.50 (d, J=17.6 Hz, 1H), 2.40 (dd, J=18.1,3.9 Hz, 1H), 2.21-2.01 (m, 4H), 1.85 (td, J=12.3, 5.5 Hz, 1H), 1.73-1.62(m, 1H), 1.29 (s, 12H).

Example A20. Preparation of4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(2-(pyridin-2-yloxy)ethyl)cyclohex-3-ene-1-carboxylicacid

Step 1. Preparation of8-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-2-oxaspiro[4.5]dec-7-en-1-one

In a 150 mL Chemglass pressure vessel with magnetic stir bar werecombined (1R,3aS,5aR,5bR,7aR,11aR,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yltrifluoromethanesulfonate (2.00 g, 2.78 mmol) with8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2-oxaspiro[4.5]dec-7-en-1-one(0.851 g, 3.06 mmol) and Buchwald precatalyst 13 (0.131 g, 0.167 mmol).The vessel was sealed with a rubber septum. A needle was inserted intothe septum and the vessel was iteratively evacuated and then purged withnitrogen in a vacuum oven at RT four times over a 15 min period. To thenitrogen purged reaction flask was added anhydrous THF (40 mL) andfreshly prepared, nitrogen sparged aqueous 0.5 M K₃PO₄ (13.9 mL, 6.95mmol) was added. The vessel was sealed and the resulting yellow solutionwas stirred at 80° C. for 20.5 h. The mixture darkened to a very deepgreen color after 30 min of heating, and after 20.5 h of heating anearly colorless biphasic mixture was present. The mixture was dilutedwith EtOAc (150 mL) and washed with saturated aqueous sodium bicarbonate(50 mL×2) and then with brine (50 mL). The combined aqueous layer wasextracted with 2×100 mL of chloroform and the organic phases werecombined, dried over anhydrous magnesium sulfate, filtered andconcentrated to a slightly yellow foam solid. The crude yellow materialwas loaded in minimum DCM onto a hexanes preequilibrated Isco 80 gsilica cartridge. Elution gradient 100% hexanes to 1:1 hexanes:EtOAcover 2 column volumes, hold 1:1 hexanes:EtOAc for 3 column volumes, thengradient 1:1 hexanes:EtOAc to 1:4 hex:EtOAc over 8 column volumes, thenhold 1:4 hexanes:EtOAc for 10 column volumes. Product fractions werecombined and concentrated in vacuo to give an off-white glassy solid:1.63 g (81.0% yield). LCMS m/z=721.6 (M+H⁺), retention time 2.404 min(LCMS Method 17). ¹H NMR (400 MHz, 1:1 mixture of CDCl3 and CD3OD, CD3ODlock) δ 5.41-5.30 (m, 1H), 5.22 (d, J=5.6 Hz, 1H), 4.70 (br. s., 1H),4.42-4.27 (m, 2H), 3.19-2.97 (m, 8H), 2.78-2.53 (m, 4H), 2.52-2.32 (m,2H), 2.29-2.10 (m, 4H), 2.04-1.75 (m, 6H), 1.69 (s, 4H), 1.66-1.54 (m,4H), 1.53 (br. s., 1H), 1.45 (br. s., 4H), 1.40-1.32 (m, 2H), 1.32-1.13(m, 5H), 1.10 (s, 6H), 1.04 (br. s., 1H), 0.99 (br. s., 5H), 0.95 (d,J=7.3 Hz, 3H), 0.88 (s, 3H).

Step 2. Preparation of potassium4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(2-hydroxyethyl)cyclohex-3-ene-1-carboxylate

In a 250 mL round bottom flask fitted with a reflux condenser werecombined8-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-2-oxaspiro[4.5]dec-7-en-1-one(1.61 g, 2.23 mmol) with potassium carbonate (1.54 g, 11.2 mmol) in amixture of MeOH (20 mL) and THF (20 mL). The result was heated to 70° C.in an oil bath for 2.5 h. Solvent was removed in vacuo to leave a solidbrown residue which was carried into the next step without furthermanipulation. LCMS m/z=739.5 (M+H⁺), retention time 1.852 min (LCMSMethod 18).

Step 3. Preparation of isopropyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(2-hydroxyethyl)cyclohex-3-ene-1-carboxylate

In a 250 mL round bottom flask fitted with a reflux condenser werecombined the crude reaction mixture from Step 2 containing potassium4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(2-hydroxyethyl)cyclohex-3-enecarboxylate(1.73 g, 2.23 mmol) with potassium carbonate (1.543 g, 11.17 mmol) in amixture of acetonitrile (20 mL) and DMF (20 mL). To the mixture wasadded 2-iodopropane (4.46 mL, 44.7 mmol). The resulting suspension wasstirred at 80° C. for 2.5 h. The mixture was concentrated in vacuo to aresidue. Ethyl acetate (120 mL) and water (100 mL) were added and themixture was shaken and phases were separated. The organic phase waswashed twice more with water (2×50 mL) and then with brine (20 mL). Theslightly yellow organic was dried over anhydrous magnesium sulfate,filtered and concentrated in vacuo to a residue. The material was loadedin DCM onto an Isco 120 g silica gel cartridge which was preequilibratedwith DCM. Elution gradient 100% DCM to 19:1 DCM MeOH over 6 columnvolumes, hold 19:1 DCM:MeOH for 8 column volumes. The combined productfractions were concentrated in vacuo to a beige foam: 1.55 g (89% yieldover 2 steps). LCMS m/z=781.5 (M+H⁺), retention time 2.873 min (LCMSMethod 19). ¹H NMR (400 MHz, CHLOROFORM-d) δ 5.34 (br. s., 1H), 5.18 (d,J=5.6 Hz, 1H), 5.04 (dt, J=12.4, 6.1 Hz, 1H), 4.73 (s, 1H), 4.61 (s,1H), 3.73 (d, J=4.9 Hz, 1H), 3.16-2.97 (m, 7H), 2.75-2.54 (m, 4H),2.54-2.42 (m, 1H), 2.28-2.16 (m, 1H), 2.13 (dd, J=12.1, 6.5 Hz, 1H),2.07-1.91 (m, 4H), 1.89-1.75 (m, 4H), 1.71 (s, 3H), 1.70-1.62 (m, 2H),1.62-1.49 (m, 5H), 1.49-1.39 (m, 4H), 1.39-1.29 (m, 3H), 1.29-1.22 (m,7H), 1.22-1.11 (m, 2H), 1.08 (s, 6H), 1.01-0.95 (m, 6H), 0.94-0.90 (m,3H), 0.88 (s, 3H).

Step 4. Preparation of isopropyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(2-((methylsulfonyl)oxy)ethyl)cyclohex-3-ene-1-carboxylate

Isopropyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(2-hydroxyethyl)cyclohex-3-enecarboxylate(0.800 g, 1.02 mmol) was dissolved in a mixture of triethylamine (5 mL)and DCM (5 mL). The clear mixture was chilled in an ice bath and to itwas slowly added a solution of methanesulfonic anhydride (0.446 g, 2.56mmol) in DCM (3 mL). The colorless solution took on a slightly yellowcolor turning to deep orange and finally to brown over the course of thereaction. The brown mixture was stirred at 0° C. for 4 h and was thenconcentrated in vacuo to a residue without warming. The crude residuewas diluted with EtOAc (100 mL) and washed with 5% aqueous NaHCO₃ (2×20mL), water (20 mL) and brine (20 mL). The organic was dried overanhydrous magnesium sulfate, filtered and concentrated in vacuo to givea reddish/brown foam. The crude material was loaded in minimum DCM ontoan 80 g Isco silica cartridge which was preequilibrated with hexanes.Elution gradient 100% hexanes to 3:2 hexanes:acetone over 3 columnvolumes, hold 3:2 hexanes:acetone for 10 column volumes. Desired productfractions were combined and concentrated in vacuo to give a yellow foam:667 mg (76.0% yield). LCMS m/z=859.6 (M+H⁺), retention time 3.160 min(LCMS Method 19). ¹H NMR (400 MHz, CHLOROFORM-d) δ 5.33 (br. s., 1H),5.18 (d, J=5.4 Hz, 1H), 5.04 (dt, J=12.2, 6.3 Hz, 1H), 4.75 (br. s.,1H), 4.63 (br. s., 1H), 4.29 (t, J=7.0 Hz, 1H), 3.72 (t, J=6.5 Hz, 1H),3.25 (s, 1H), 3.16 (s, 1H), 3.08 (br. s., 6H), 3.01 (s, 2H), 2.83 (s,1H), 2.77-2.54 (m, 4H), 2.49 (br. s., 1H), 2.30-2.09 (m, 3H), 2.09-1.95(m, 4H), 1.95-1.76 (m, 4H), 1.72 (br. s., 3H), 1.66 (dd, J=14.3, 7.2 Hz,3H), 1.61-1.50 (m, 5H), 1.50-1.38 (m, 5H), 1.33 (t, J=13.1 Hz, 3H),1.29-1.21 (m, 7H), 1.18-1.03 (m, 6H), 1.00 (br. s., 3H), 0.97 (d, J=7.3Hz, 3H), 0.93 (d, J=5.4 Hz, 3H), 0.88 (s, 3H).

Step 5. Preparation of isopropyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(2-(pyridin-2-yloxy)ethyl)cyclohex-3-ene-1-carboxylate

In a 1 dram vial with PTFE screwcap were combined pyridin-2-ol (0.0190g, 0.204 mmol) and isopropyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(2-((methylsulfonyl)oxy)ethyl)cyclohex-3-enecarboxylate(0.0250 g, 0.0290 mmol) in anhydrous DMF (0.5 mL). To the mixture wasadded NaHMDS, 1.0M in THF (0.175 mL, 0.175 mmol) with stirring. Theresulting slightly yellow mixture was heated to 50° C. and stirred for 3d. The crude mixture was purified by reverse phase preparative HPLC(Preparative HPLC Method 6). Thus was isolated the desired material(0.00940 g, 29.7% yield) as a white solid TFA salt. LCMS m/z=858.6(M+H⁺), retention time 1.627 min (LCMS Method 16).

Step 6. In a 1 dram vial with PTFE screwcap were combined isopropyl4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(2-(pyridin-2-yloxy)ethyl)cyclohex-3-enecarboxylate,TFA salt (0.00940 g, 8.65 μmol) with lithium hydroxide, 1.0M aqueous(0.087 mL, 0.087 mmol) and a mixture of THF (0.3 mL) and MeOH (0.3 mL).The resulting mixture was stirred at 75° C. for 48 h. The crude mixturewas purified by reverse phase preparative HPLC (Preparative HPLC Method6). The fraction containing the desired material was concentrated invacuo to give the title compound as a white glassy solid (0.0035 g 33%yield). LCMS m/z=816.5 (M+H⁺), retention time 2.182 min (LCMS Method17). ¹H NMR (400 MHz, 1:1 mixture of CDCl3 and CD3OD, CD3OD lock) δ 8.07(d, J=5.1 Hz, 1H), 7.65-7.59 (m, 1H), 6.90 (t, J=6.1 Hz, 1H), 6.74 (d,J=8.3 Hz, 1H), 5.33 (br. s., 1H), 5.18 (d, J=5.6 Hz, 1H), 4.80 (s, 1H),4.72 (s, 1H), 4.34 (t, J=6.6 Hz, 2H), 3.24 (br. s., 3H), 3.21-3.13 (m,3H), 3.12-2.96 (m, 4H), 2.84-2.72 (m, 1H), 2.60 (d, J=15.4 Hz, 1H),2.26-1.96 (m, 10H), 1.87-1.70 (m, 6H), 1.69-1.59 (m, 3H), 1.57 (br. s.,2H), 1.53-1.43 (m, 5H), 1.40 (br. s., 1H), 1.39-1.22 (m, 2H), 1.15 (s,3H), 1.11 (br. s., 1H), 1.08 (s, 3H), 1.04-0.99 (m, 1H), 0.97 (br. s.,3H), 0.93 (s, 3H), 0.90 (s, 3H).

Example A21. Preparation of4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(2-((5-methylisothiazol-3-yl)oxy)ethyl)cyclohex-3-ene-1-carboxylicacid

The title compound was obtained by the same procedures used in thepreparation of4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(2-(pyridin-2-yloxy)ethyl)cyclohex-3-ene-1-carboxylicacid, except 5-methylisothiazol-3-ol (0.023 g, 0.204 mmol) was used inplace of pyridin-2-ol in Step 5. Thus was obtained the title compound asa white glassy solid (0.0027 g, 8.3% combined yield for Steps 5 and 6).LCMS m/z=836.5 (M+H⁺), retention time 2.394 min (LCMS Method 17). ¹HNMR(400 MHz, 1:1 mixture of CDCl3 and CD3OD, CD3OD lock) δ 5.32 (br. s.,1H), 5.18 (d, J=5.4 Hz, 1H), 4.80 (s, 1H), 4.73 (s, 1H), 4.36 (t, J=6.6Hz, 2H), 3.75 (t, J=6.0 Hz, 2H), 3.27-3.12 (m, 8H), 3.12-2.94 (m, 5H),2.78 (td, J=10.8, 4.4 Hz, 1H), 2.58 (d, J=16.1 Hz, 1H), 2.25-1.95 (m,11H), 1.92-1.70 (m, 8H), 1.70-1.59 (m, 3H), 1.59-1.39 (m, 9H), 1.39-1.24(m, 3H), 1.22 (s, 1H), 1.18-1.04 (m, 7H), 0.97 (d, J=2.7 Hz, 3H), 0.92(br. s., 3H), 0.90 (s, 3H).

Example A22. Preparation of1-(2-((3-cyanopyridin-2-yl)oxy)ethyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylicacid

Step 1. Preparation of potassium4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(2-hydroxyethyl)cyclohex-3-ene-1-carboxylate

In a 50 mL round bottom flask fitted with a reflux condenser werecombined8-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-2-oxaspiro[4.5]dec-7-en-1-one(0.700 g, 0.971 mmol) with potassium carbonate (1.34 g, 9.71 mmol) in amixture of MeOH (10 mL) and THF (15 mL). The result was heated to refluxin an 85° C. oil bath for 24 h. The mixture was allowed to cool to RT,then DCM was added and the result was filtered to remove white solids.Solvent was removed in vacuo and the residue was dried in a vacuum ovenat 50° C. overnight to afford the desired material as a white powder(0.940 g, 125% yield). Mass recovery indicated that the material wasapproximately 80% pure with the remainder as excess potassium salts.This material was used directly in the next step without furtherpurification. LCMS m/z=739.5 (M+H⁺), retention time 1.852 min (LCMSMethod 17).

Step 2. To the crude powder product from Step 1 containing approx. 80%by weight potassium4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(2-hydroxyethyl)cyclohex-3-enecarboxylate(0.025 g, 0.026 mmol) was added 2-fluoronicotinonitrile (0.016 g, 0.129mmol), anhydrous DMF (0.4 mL) and anhydrous THF (0.3 mL) to give aslightly cloudy yellow mixture. To the mixture was added potassiumtert-butoxide, 1.0M in THF (0.103 mL, 0.103 mmol). The mixture wasstirred at RT for 2 h, and then additional 6-fluoropicolinonitrile(0.032 g, 0.258 mmol) and potassium tert-butoxide, 1.0M in THF (0.206mL, 0.206 mmol) and more DMF (0.2 mL) were added and the mixture wasstirred for another 1 h. The crude mixture was purified by reverse phasepreparative HPLC (Preparative HPLC Method 7). The title compound wasthus obtained as a slightly yellow powder (0.0086 g, 25% yield) as a TFAsalt. LCMS m/z=841.6 (M+H⁺), retention time 2.289 min (LCMS Method 17).H NMR (400 MHz, 1:1 mixture of CDCl3 and CD3OD, CD3OD lock) δ 8.34 (dd,J=4.2, 1.0 Hz, 1H), 7.94 (dd, J=8.1, 1.7 Hz, 1H), 7.03 (dd, J=7.1, 5.1Hz, 1H), 5.40-5.28 (m, 1H), 5.17 (d, J=4.6 Hz, 1H), 4.80 (br. s., 1H),4.71 (br. s., 1H), 4.39-4.31 (m, 1H), 3.28-3.12 (m, 7H), 3.09 (br. s.,2H), 3.01 (br. s., 2H), 2.82 (br. s., 1H), 2.61 (d, J=17.4 Hz, 1H),2.25-1.96 (m, 10H), 1.86 (d, J=10.5 Hz, 1H), 1.78-1.68 (m, 5H),1.67-1.53 (m, 5H), 1.53-1.38 (m, 6H), 1.38-1.24 (m, 3H), 1.19-1.03 (m,8H), 1.03-0.82 (m, 9H).

Example A23. Preparation of2-(2-(1-carboxy-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-en-1-yl)ethoxy)isonicotinicacid

Step 1. Preparation of potassium4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(2-hydroxyethyl)cyclohex-3-ene-1-carboxylate

In a 50 mL round bottom flask fitted with a reflux condenser werecombined8-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-2-oxaspiro[4.5]dec-7-en-1-one(0.700 g, 0.971 mmol) with potassium carbonate (1.34 g, 9.71 mmol) in amixture of MeOH (10 mL) and THF (15 mL). The result was heated to refluxin an 85° C. oil bath for 24 h. The mixture was allowed to cool to RT,then DCM was added and the result was filtered to remove white solids.Solvent was removed in vacuo and the residue was dried in a vacuum ovenat 50° C. overnight to afford the desired material as a white powder(0.940 g, 125% yield). 0.9155 g of this material was dissolved withstirring in 10 mL of 9:1 DCM:MeOH and this suspension (salts did notdissolve) was loaded onto a short 40 mL silica gel plug in a 60 mL glassfrit suction funnel. The material was eluted with 400 mL of 9:1DCM:MeOH. Much of the orange color associated with the impure productwas left behind on the silica. Concentration in vacuo afforded apinkish/white solid which was placed in a vacuum oven at 45° C. forseveral hours. The desired material was thus obtained as a white powder(0.5082 g, 69.4% yield). LCMS m/z=739.6 (M+H⁺), retention time 1.978 min(LCMS Method 21).

Step 2. To the purified Step 1 product potassium4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(2-hydroxyethyl)cyclohex-3-enecarboxylate(0.025 g, 0.032 mmol) was added 2-chloroisonicotinic acid (0.025 g,0.161 mmol) followed by anhydrous DMF (0.35 mL). To the mixture wasadded potassium tert-butoxide, 1.0M in THF (0.322 mL, 0.322 mmol). Themixture became slightly yellow and cloudy with suspended solid uponaddition of the base. The mixture was stirred at RT for 70 h. Thereaction mixture was quenched by addition of 3 drops of acetic acid. 0.5mL MeOH was then added and the mixture was filtered. The crude mixturewas purified by reverse phase preparative HPLC in a single injection(Preparative HPLC Method 8). Thus was obtained the title compound as awhite solid (0.0069 g, 18% yield) TFA salt. LCMS m/z=860.6 (M+H⁺),retention time 1.559 min (LCMS Method 20).

Example A24. Preparation of1-(2-((4-cyanopyridin-2-yl)oxy)ethyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylicacid

The title compound was prepared following the procedure described forthe preparation of2-(2-(1-carboxy-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-en-1-yl)ethoxy)isonicotinicacid, except in the present case 2-fluoroisonicotinonitrile (0.020 g,0.161 mmol) was used instead of 2-chloroisonicotinic acid, and there wasalso less potassium tert-butoxide, 1.0M in THF used in the present case(0.129 mL, 0.129 mmol). The title compound was isolated as a slightlyyellow solid (0.0133 g, 36.0% yield) TFA salt. LCMS m/z=841.6 (M+H⁺),retention time 1.689 min (LCMS Method 20). ¹H NMR (400 MHz, 1:1 mixtureof CDCl3 and CD3OD, CD3OD lock) δ 8.28 (d, J=5.4 Hz, 1H), 7.11 (dd,J=5.1, 1.2 Hz, 1H), 7.00 (s, 1H), 5.33 (br. s., 1H), 5.17 (d, J=4.4 Hz,1H), 4.79 (s, 1H), 4.71 (s, 1H), 4.47-4.39 (m, 2H), 3.28-3.04 (m, 9H),3.04-2.96 (m, 2H), 2.86-2.74 (m, 1H), 2.59 (d, J=16.4 Hz, 1H), 2.24-1.95(m, 11H), 1.89-1.74 (m, 3H), 1.73 (s, 4H), 1.68-1.42 (m, 10H), 1.42-1.29(m, 3H), 1.15 (s, 3H), 1.11 (br. s., 2H), 1.08 (s, 4H), 0.96 (d, J=2.4Hz, 3H), 0.92 (d, J=2.9 Hz, 3H), 0.90 (s, 3H).

Example A25. Preparation of4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(2-(pyrimidin-2-yloxy)ethyl)cyclohex-3-ene-1-carboxylicacid

The title compound was prepared following the procedure described forthe preparation of2-(2-(1-carboxy-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-en-1-yl)ethoxy)isonicotinicacid, except in the present case 2-bromopyrimidine (0.026 g, 0.161 mmol)was used instead of 2-chloroisonicotinic acid, and there was also lesspotassium tert-butoxide, 1.0M in THF used in the present case (0.129 mL,0.129 mmol). The title compound was isolated as a white solid (0.0056 g,14.2% yield) TFA salt. LCMS m/z=817.6 (M+H⁺), retention time 1.547 min(LCMS Method 20). ¹H NMR (400 MHz, 1:1 mixture of CDCl3 and CD3OD, CD3ODlock) δ 8.50 (d, J=4.9 Hz, 1H), 7.02 (t, J=4.8 Hz, 1H), 5.35 (dd, J=4.1,2.9 Hz, 1H), 5.25-5.14 (m, 1H), 4.80 (s, 1H), 4.72 (s, 1H), 4.50-4.44(m, 1H), 4.38-4.31 (m, 1H), 3.27-2.98 (m, 10H), 2.84-2.75 (m, 1H),2.64-2.58 (m, 1H), 2.25-1.96 (m, 10H), 1.89-1.75 (m, 3H), 1.73 (s, 3H),1.69-1.53 (m, 5H), 1.53-1.25 (m, 8H), 1.15 (d, J=2.9 Hz, 3H), 1.11 (br.s., 2H), 1.08 (s, 3H), 1.03-0.84 (m, 9H).

Example A26. Preparation of4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(2-((4-methylpyrimidin-2-yl)oxy)ethyl)cyclohex-3-ene-1-carboxylicacid

The title compound was prepared following the procedure described forthe preparation of2-(2-(1-carboxy-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-en-1-yl)ethoxy)isonicotinicacid, except in the present case 2-chloro-4-methylpyrimidine (0.021 g,0.161 mmol) was used instead of 2-chloroisonicotinic acid, and there wasalso less potassium tert-butoxide, 1,0M in THF used in the present case(0.129 mL, 0.129 mmol). The title compound was isolated as a white solid(0.0056 g, 14.2% yield) TFA salt. LCMS m/z=831.7 (M+H⁺), retention time1.550 min (LCMS Method 20). ¹H NMR (400 MHz, 1:1 mixture of CDCl3 andCD3OD, CD3OD lock) δ 8.21 (d, J=6.1 Hz, 1H), 6.52 (d, J=5.9 Hz, 1H),5.40-5.34 (m, 1H), 5.23 (d, J=4.6 Hz, 1H), 4.79 (s, 1H), 4.71 (s, 1H),4.41-4.27 (m, 2H), 3.30-3.05 (m, 10H), 3.01 (d, <7=3.4 Hz, 2H), 2.81(td, J=11.2, 4.9 Hz, 1H), 2.49-2.33 (m, 2H), 2.27-1.98 (m, 10H),1.93-1.81 (m, 2H), 1.81-1.74 (m, 2H), 1.72 (s, 4H), 1.69-1.40 (m, 12H),1.38-1.34 (m, 1H), 1.21-1.03 (m, 9H), 1.02-0.86 (m, 8H).

Example A27. Preparation of4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(2-((4-methoxypyrimidin-2-yl)oxy)ethyl)cyclohex-3-ene-1-carboxylicacid

The title compound was prepared following the procedure described forthe preparation of2-(2-(1-carboxy-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-en-1-yl)ethoxy)isonicotinicacid, except in the present case 2-chloro-4-methoxypyrimidine (0.023 g,0.161 mmol) was used instead of 2-chloroisonicotinic acid, and there wasalso less potassium tert-butoxide, 1.0M in THF used in the present case(0.129 mL, 0.129 mmol). The title compound was isolated as a white solid(0.0116 g, 28.8% yield) TFA salt. LCMS m/z=847.7 (M+H⁺), retention time1.525 min (LCMS Method 20). ¹H NMR (400 Mhz, 1:1 mixture of CDCl3 andCD3OD, CD3OD lock) δ 7.28 (d, J=7.6 Hz, 1H), 5.62 (d, J=7.6 Hz, 1H),5.40-5.33 (m, 1H), 5.23 (d, J=4.9 Hz, 1H), 4.79 (s, 1H), 4.71 (s, 1H),4.41-4.29 (m, 2H), 3.27-2.97 (m, 12H), 2.81 (td, J=11.2, 4.9 Hz, 1H),2.43-2.33 (m, 1H), 2.28-1.98 (m, 10H), 1.92-1.81 (m, 2H), 1.80-1.73 (m,2H), 1.73 (s, 3H), 1.70-1.40 (m, 12H), 1.38-1.34 (m, 1H), 1.16 (s, 3H),1.15-1.09 (m, 2H), 1.08 (s, 3H), 1.00 (d, J=3.2 Hz, 3H), 0.96 (d, J=7.6Hz, 3H), 0.91 (s, 3H).

Example A28. Preparation of4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(2-((3-methylpyridin-2-yl)oxy)ethyl)cyclohex-3-ene-1-carboxylicacid

The title compound was prepared following the procedure described forthe preparation of2-(2-(1-carboxy-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-en-1-yl)ethoxy)isonicotinicacid, except in the present case 2-fluoro-3-methylpyridine (0.018 g,0.161 mmol) was used instead of 2-chloroisonicotinic acid, and there wasalso less potassium tert-butoxide, 1.0M in THF used in the present case(0.129 mL, 0.129 mmol). The title compound was isolated as a white solid(0.0262 g, 74.7% yield) TFA salt. LCMS m/z=830.7 (M+F1⁺), retention time1.707 min (LCMS Method 20). NMR (400 MHz, 1:1 mixture of CDCl3 andCD3OD, CD3OD lock) δ 7.89 (dd, J=5.1, 1.2 Hz, 0.35H), 7.44 (dd, J=7.1,1.0 Hz, 0.35H), 7.42-7.37 (m, 0.65H), 7.22 (dd, J=6.5, 1.3 Hz, 0.65H),6.81 (dd, J=7.0, 5.3 Hz, 0.35H), 6.29 (t, J=6.1 Hz, 0.65H), 5.39-5.30(m, 1H), 5.23 (d, J=4.9 Hz, 0.65H), 5.18 (d, J=4.6 Hz, 0.35H), 4.79 (s,1H), 4.71 (s, 1H), 4.41-4.29 (m, 2H), 3.27-2.98 (m, 12H), 2.81 (td,J=11.1, 4.6 Hz, 1H), 2.43-2.33 (m, 1H), 2.30-2.07 (m, 10H), 2.07-1.94(m, 4H), 1.92-1.73 (m, 4H), 1.72 (s, 3H), 1.69-1.40 (m, 12H), 1.38-1.34(m, 1H), 1.20-1.05 (m, 9H), 1.02-0.86 (m, 9H).

Example A29. Preparation of1-(2-((3-chloropyridin-2-yl)oxy)ethyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylicacid

The title compound was prepared following the procedure described forthe preparation of2-(2-(1-carboxy-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-en-1-yl)ethoxy)isonicotinicacid, except in the present case 3-chloro-2-fluoropyridine (0.021 g,0.161 mmol) was used instead of 2-chloroisonicotinic acid, and there wasalso less potassium tert-butoxide, 1.0M in THF used in the present case(0.129 mL, 0.129 mmol). The title compound was isolated as a white solid(0.0156 g, 42.7% yield) TFA salt. LCMS m/z=850.6 (M+H⁺), retention time1.770 min (LCMS Method 20). ¹H NMR (400 MHz, 1:1 mixture of CDCl3 andCD3OD, CD3OD lock) δ 7.99 (dd, J=4.9, 1.7 Hz, 1H), 7.65 (dd, J=7.7, 1.6Hz, 1H), 6.86 (dd, J=7.6, 4.9 Hz, 1H), 5.33 (br. s., 1H), 5.17 (d, J=4.6Hz, 1H), 4.79 (s, 1H), 4.72 (s, 1H), 4.45 (t, J=6.6 Hz, 2H), 3.27-2.98(m, 12H), 2.80 (td, J=11.1, 4.8 Hz, 1H), 2.60 (d, J=15.7 Hz, 1H),2.25-1.95 (m, 10H), 1.90-1.74 (m, 3H), 1.73 (s, 3H), 1.68-1.42 (m, 10H),1.40 (br. s., 1H), 1.38-1.29 (m, 2H), 1.29-1.23 (m, 1H), 1.15 (s, 3H),1.12 (d, J=5.4 Hz, 1H), 1.08 (s, 3H), 0.99-0.84 (m, 9H).

Example A30. Preparation of4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(2-((3-(trifluoromethyl)pyridin-2-yl)oxy)ethyl)cyclohex-3-ene-1-carboxylicacid

The title compound was prepared following the procedure described forthe preparation of2-(2-(1-carboxy-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-en-1-yl)ethoxy)isonicotinicacid, except in the present case 2-chloro-3-(trifluoromethyl)pyridine(0.029 g, 0.161 mmol) was used instead of 2-chloroisonicotinic acid, andthere was also less potassium tert-butoxide, 1.0M in THF used in thepresent case (0.129 mL, 0.129 mmol). The title compound was isolated asa white solid (0.0020 g, 4.9% yield) TFA salt. LCMS m/z=884.6 (M+H⁺),retention time 1.810 min (LCMS Method 20).

Example A31. Preparation of4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(2-(pyrazin-2-yloxy)ethyl)cyclohex-3-ene-1-carboxylicacid

The title compound was prepared following the procedure described forthe preparation of2-(2-(1-carboxy-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-en-1-yl)ethoxy)isonicotinicacid, except in the present case 2-chloropyrazine (0.018 g, 0.161 mmol)was used instead of 2-chloroisonicotinic acid, and there was also lesspotassium tert-butoxide, 1.0M in THF used in the present case (0.129 mL,0.129 mmol). The title compound was isolated as a white solid (0.0102 g,28.2% yield) TFA salt. LCMS m/z=817.6 (M+H⁺), retention time 1.592 min(LCMS Method 20). ¹H NMR (400 MHz, 1.1 mixture of CDCl3 and CD3OD, CD3ODlock) δ 8.16-8.08 (m, 2H), 8.05 (d, J=2.4 Hz, 1H), 5.33 (br. s., 1H),5.18 (d, J=5.4 Hz, 1H), 4.79 (s, 1H), 4.71 (br. s., 1H), 4.44 (t, J=6.2Hz, 2H), 3.27-3.13 (m, 7H), 3.13-3.05 (m, 3H), 3.05-2.95 (m, 2H),2.86-2.74 (m, 1H), 2.60 (d, J=17.4 Hz, 1H), 2.25-1.96 (m, 10H),1.89-1.81 (m, 1H), 1.81-1.74 (m, 2H), 1.73 (s, 4H), 1.65-1.42 (m, 10H),1.40 (br. s., 1H), 1.38-1.24 (m, 3H), 1.15 (s, 3H), 1.11 (br. s., 2H),1.08 (s, 3H), 1.01-0.86 (m, 9H).

Example A32. Preparation of4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)-1-(2-((4-methoxypyridin-2-yl)oxy)ethyl)cyclohex-3-ene-1-carboxylicacid

The title compound was prepared following the procedure described forthe preparation of2-(2-(1-carboxy-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-en-1-yl)ethoxy)isonicotinicacid, except in the present case 2-bromo-4-methoxypyridine (0.030 g,0.161 mmol) was used instead of 2-chloroisonicotinic acid, and there wasalso less potassium tert-butoxide, 1.0M in THF used in the present case(0.129 mL, 0.129 mmol). The title compound was one of two compoundsisolated from this reaction. The material was obtained as a white solid(0.0068 g, 18.3% yield) TFA salt. LCMS m/z=846.7 (M+H⁺), retention time1.335 min (LCMS Method 20). ¹H NMR (400 MHz, 1:1 mixture of CDCl3 andCD3OD, CD3OD lock) δ 7.97 (d, 2=6.6 Hz, 1H), 6.76 (dd, 2=6.6, 2.2 Hz,1H), 6.59 (d, 2=2.0 Hz, 1H), 5.34 (br. s., 1H), 5.18 (d, 2=4.6 Hz, 1H),4.79 (s, 1H), 4.71 (s, 1H), 4.42 (t, 2=6.7 Hz, 2H), 3.98 (s, 3H),3.27-3.04 (m, 10H), 3.01 (d, 2=3.4 Hz, 2H), 2.86-2.76 (m, 1H), 2.67-2.57(m, 1H), 2.27-2.15 (m, 3H), 2.15-1.96 (m, 8H), 1.85 (td, 2=12.2, 3.3 Hz,1H), 1.81-1.73 (m, 2H), 1.72 (s, 4H), 1.66-1.38 (m, 10H), 1.38-1.28 (m,2H), 1.15 (s, 3H), 1.12 (br. s., 2H), 1.07 (s, 3H), 1.01-0.85 (m, 9H).

Example A33. Preparation of1-(2-((4-bromopyridin-2-yl)oxy)ethyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylicacid

The title compound was prepared following the procedure described forthe preparation of2-(2-(1-carboxy-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-en-1-yl)ethoxy)isonicotinicacid, except in the present case 2-bromo-4-methoxypyridine (0.030 g,0.161 mmol) was used instead of 2-chloroisonicotinic acid, and there wasalso less potassium tert-butoxide, 1.0M in THF used in the present case(0.129 mL, 0.129 mmol). The title compound was one of two compoundsisolated from this reaction. The material was obtained as a white solid(0.0045 g, 12.2% yield) TFA salt. LCMS m/z=894.5 (M+F1⁺), retention time1.672 min (LCMS Method 20). ¹H NMR (400 MHz, 1:1 mixture of CDCl3 andCD3OD, CD3OD lock) δ 8.09 (d, J=5.9 Hz, 1H), 7.06 (d, J=2.2 Hz, 1H),6.86 (dd, J=5.9, 2.2 Hz, 1H), 5.33 (br. s., 1H), 5.21-5.15 (m, 1H), 4.80(s, 1H), 4.72 (s, 1H), 4.16 (t, J=6.6 Hz, 2H), 3.27-2.98 (m, 12H),2.84-2.74 (m, 1H), 2.60 (dd, J=18.7, 2.8 Hz, 1H), 2.24-1.96 (m, 11H),1.87-1.74 (m, 3H), 1.73 (s, 4H), 1.68-1.55 (m, 4H), 1.55-1.38 (m, 7H),1.38-1.25 (m, 2H), 1.15 (s, 3H), 1.14-1.10 (m, 1H), 1.08 (s, 3H),1.01-0.96 (m, 3H), 0.96-0.91 (m, 3H), 0.90 (s, 3H).

Example A34. Preparation of1-(2-((4-chloropyridin-2-yl)oxy)ethyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylicacid

The title compound was prepared following the procedure described forthe preparation of2-(2-(1-carboxy-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-en-1-yl)ethoxy)isonicotinicacid, except in the present case 2,4-dichloropyridine (0.024 g, 0.161mmol) was used instead of 2-chloroisonicotinic acid, and there was alsoless potassium tert-butoxide, 1.0M in THF used in the present case(0.129 mL, 0.129 mmol). The title compound was one of two compoundsisolated from this reaction. The material was obtained as a slightlyyellow solid (0.0143 g, 38.7% yield) TFA salt. LCMS m/z=850.6 (M+H⁺),retention time 1.637 min (LCMS Method 20). ¹H NMR (400 MHz, 1:1 mixtureof CDCl3 and CD3OD, CD3OD lock) δ 8.11 (d, J=5.9 Hz, 1H), 6.91 (d, J=2.2Hz, 1H), 6.83 (dd, J=5.9, 2.2 Hz, 1H), 5.33 (br. s., 1H), 5.18 (d, J=4.6Hz, 1H), 4.79 (s, 1H), 4.71 (s, 1H), 4.17 (t, J=6.6 Hz, 2H), 3.27-2.98(m, 12H), 2.80 (td, J=11.2, 4.8 Hz, 1H), 2.60 (d, J=16.6 Hz, 1H),2.26-1.97 (m, 11H), 1.89-1.74 (m, 3H), 1.72 (s, 4H), 1.67-1.38 (m, 11H),1.38-1.27 (m, 2H), 1.15 (s, 3H), 1.10 (d, J=11.0 Hz, 1H), 1.07 (s, 3H),0.99-0.86 (m, 9H).

Example A35. Preparation of1-(2-((2-chloropyridin-4-yl)oxy)ethyl)-4-((1R,3aS,5aR,5bR,7aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-ene-1-carboxylicacid

The title compound was prepared following the procedure described forthe preparation of 2-(2-(1-carboxy-4-((1R,3aS,5 aR,5bR,7 aR,11aS,11bR,13aR,13bR)-3a-((2-(1,1-dioxidothiomorpholino)ethyl)amino)-5a,5b,8,8,11a-pentamethyl-1-(prop-1-en-2-yl)-2,3,3a,4,5,5a,5b,6,7,7a,8,11,11a,11b,12,13,13a,13b-octadecahydro-1H-cyclopenta[a]chrysen-9-yl)cyclohex-3-en-1-yl)ethoxy)isonicotinicacid, except in the present case 2,4-dichloropyridine (0.024 g, 0.161mmol) was used instead of 2-chloroisonicotinic acid, and there was alsoless potassium tert-butoxide, 1.0M in THF used in the present case(0.129 mL, 0.129 mmol). The title compound was one of two compoundsisolated from this reaction. The material was obtained as a slightlyyellow solid (0.0168 g, 41.2% yield) TFA salt. LCMS m/z=850.6 (M+H⁺),retention time 1.809 min (LCMS Method 20). ¹H NMR (400 MHz, 1:1 mixtureof CDCl3 and CD3OD, CD3OD lock) δ 8.00 (d, J=5.6 Hz, 1H), 6.90 (dd,J=5.6, 1.7 Hz, 1H), 6.76 (d, J=1.7 Hz, 1H), 5.32 (br. s., 1H), 5.17 (d,J=4.9 Hz, 1H), 4.79 (s, 1H), 4.71 (s, 1H), 4.41-4.33 (m, 2H), 3.28-2.98(m, 12H), 2.80 (td, J=1.0, 4.6 Hz, 1H), 2.58 (d, J=15.4 Hz, 1H),2.26-1.96 (m, 11H), 1.89-1.73 (m, 2H), 1.72 (s, 4H), 1.67-1.38 (m, 11H),1.38-1.26 (m, 2H), 1.18-1.04 (m, 8H), 1.01-0.86 (m, 9H).

HIV Cell Culture Assay

Cells. MT-2 cells and 293T cells were obtained from the NIH AIDSResearch and Reference Reagent Program. Cell lines were sub-culturedtwice a week in either RPMI 1640 (MT-2) or DMEM (293T, HeLa) mediasupplemented with 10% heat inactivated fetal bovine serum (FBS), 100units/mL of penicillin G and 100 μg/mL of streptomycin. The DMEM mediumwas additionally supplemented with 10 mM HEPES buffer, pH 7.55, 2 mML-glutamine and 0.25 μg/mL of amphotericin B.

Viruses. NLRepRluc virus contains the Renilla luciferase marker in placeof the viral nef gene. The proviral plasmid pNLRepRluc was constructedat Bristol-Myers Squibb, starting from a proviral NL4-3 clone (Bsubtype) that was obtained from the NIH AIDS Research and ReferenceReagent Program. The parental recombinant wild type (WT) virus(NLRepRlucP373S) was derived from NLRepRluc and contains the additionalsubstitution of P373 for serine in Gag (within the SP1 spacer), the mostcommon 373 variation in subtype B. Other recombinant viruses (A364V,V370A/AT371 and the “T332S triple” (T332S/V362I+HIV-1 protease R41G))were generated by site-directed mutagenesis of plasmid pNLRepRlucP373Sto introduce those amino acid substitutions in Gag and protease.Recombinant virus DNA was then used to generate virus stocks bytransfection of 293T cells (Lipofectamine PLUS kit, Invitrogen). Titersof virus stocks were determined using a luciferase assay(Dual-Luciferase® Reporter Assay System, Promega, Milwaukee, Wis., USA)endpoint.

Multiple cycle drug susceptibility assay. Pellets of MT-2 cells wereinfected with NLRepRlucP373S Gag site-directed viruses, where initialinocula of the reporter strains were normalized using equivalentendpoint luciferase activity signals. Such cell-virus mixtures wereresuspended in medium, incubated for 1-hour at 37° C./CO₂, and added tocompound containing 96-well plates at a final cell density of 10,000cells per well. The test compounds were 3-fold serially diluted in 100%DMSO, and assayed at a final DMSO concentration of 1%. After 4-5 dayincubation at 37° C./CO₂, virus yields were determined by Renillaluciferase activity (Dual-Luciferase® Reporter Assay System, Promega).The endpoint luminescence was detected on a Wallac Trilux (PerkinElmer).

The 50% inhibitory concentrations (EC₅₀s) were calculated by using theexponential form of the median effect equation where PercentInhibition=1/[1+(EC₅₀/drug cone.)], where m is a parameter that reflectsthe slope of the concentration-response curve. Background was taken asthe residual signal observed upon inhibition at the highestconcentration of a control protease inhibitor, NFV (3 μM).

The 90% inhibitory concentrations (EC₉₀s) were calculated by using theexponential form of the median effect equation whereEC_(F)=[(F/(100−F)]^(1/H)·EC₅₀, where H is a parameter that reflects theslope of the concentration-response curve. Background was taken as theresidual signal observed upon inhibition at the highest concentration ofa control protease inhibitor, NFV (3 μM).

HIV Cell Culture Assay

HIV-1 NL₄₋₃ expressing Renilla luciferase gene was converted to the gagV370A/ΔT371 virus by site directed mutagenesis. A364V is a site directedmutant.

T332s/V362I/Pr R41G (N14.3, B Clade) virus' was obtained as follows:Selection for resistance of HIV-1 strain NL4-4 virus with the HIVmaturation inhibitor (MI) compound

was started at the EC₅₀ for this virus (2 nM), with a two-fold increasein the maturation inhibitor compound concentration applied at eachpassage. At passage 8 virus was harvested and sequenced. The selectedvirus population contained Gag amino acid substitutions T332S and V362Iand the R41G substitution in protease. These substitutions weresubsequently introduced into NLRepRlucP373, a derivative of HIV-1 cloneNL4-3 modified to contain P373S, the most common polymorphicsubstitution in subtype B at position 373, and the Renilla luciferasegene inserted into the ne/locus.

The emergence of selected substitutions in the wt genotypic backgroundis discussed herein:

Starting from wt virus, the HIV protease R41G substitution was detectedin one of three in vitro selections for resistance to the MI compoundabove along with Gag V362I and Gag T332S. R41G is not a primary PIresistance substitution^(i) and is not present in the LANL database(2010). There is a single report of R41G associated with in vitroselection for resistance to an investigational PI^(ii). However, in thatcase, R41G did not itself convey PI resistance. A related change, R41K,is a common subtype B polymorph (27% of LANL database), and R41K may beinvolved in the emergence of protease resistance to an investigationalprotease inhibitor.^(iii) R41 is located in a loop proximal to the HIV-1protease substrate binding site, and this change might actallosterically to facilitate closing the protease active site pocketover the substrate, thereby allowing catalysis. It might be that R41Galters the dynamics of the loop motion and the final positioning of theloop, which could cause the active site to better recognize the primaryMI compound (above)-selected changes (V362I/T332S). An analysis of theV362I/T332S/Pr R41G substitutions, and their effects on MI compoundsusceptibility and viral growth, are described in the Table 1 below:

TABLE 1 Anti-Viral Sensitivity of Site Directed Mutants Virus titer,TCID₅₀ Fold wt (×10⁵/mL) MI BVM Group Genotype CPE Rluc RT Compound(Bevirimat) Key substitutions Crosswise effects of T332S and Pr R41G onV362I 6 V362I 2.6 1.6 2.6 2.2 0.6 T332S 2.6 6.6 0.4 1.9 23 HIV protease2.6 2.6 1.0 1.5 1.9 R41G T332S/V362I/ 4.1 6.6 4.1 5.7 3.1 T332S/prR41G0.6 1.0 0.4 6.1 4.2 V362I/prR41G 0.6 1.6 0.6 9.3 3.9 T332S/V362I/ 0.31.6 0.1 217 10 Pr41G

Viruses were constructed that contain T332S and HIV protease R41Gcombinations, with and without V362I.). Viruses with only a singlechange are only ˜2-fold less sensitive to the MI compound, while doublecombinations of these 3 substitutions are 5.7- to 9.3-fold lesssensitive. The virus with the triple change is much less sensitive tothe MI compound, suggesting that the R41G change in protease may‘crosstalk’ with Gag changes to further reduce sensitivity to the MIcompound, an unexpected finding. Thus, the T332S/V362I Site directedmutant (SDM) virus exhibits a fold change of only 5.7, but addition ofthe R41G protease change substantially increases the FC to 217.

-   ^(i) Johnson V A, Brun-Vezinet F, Clotet B, Gunthard H F, Kuritzkes    D R, Pillay D, Schapiro J M, Richman D D. Update of the drug    resistance mutations in HIV-1: December 2009. Top HIV Med. 2009    December; 17(5): 138-45.-   ^(ii) Dierynck, I, Van Markck, H, Van Ginderen, M, Jonckers, T H,    Nalam, M N, Schiffer, C A, Raoof, A, Kraus, G, Picchio, G.    TMC310911, a novel human immunodeficiency virus type 1 protease    inhibitor, shows in vitro an improved resistance profile and higher    genetic barrier to resistance compared-   ^(iii) Stray K M, Callebaut C, Glass B, Tsai L, Xu L, Müller B,    Kräusslich H G, Cihlar T. Mutations in multiple domains of Gag drive    the emergence of in vitro resistance to the phosphonate-containing    HIV-1 protease inhibitor GS-8374. J Virol. 2013 87:454-63

All three recombinant viruses were used as described above in the HIVcell culture assay for the NL₄₋₃ virus. The EC₅₀WT, EC₅₀V370A/ΔT371,EC₅₀ A364V and EC₅₀ T332s/V362I/Pr R41G data for the compounds is shownin Table 2.

Biological Data Key for EC_(50S) Compounds with Compounds with EC₅₀ >0.05 μM EC₅₀ < 0.0.5 μM Group “B” Group “A”

TABLE 2 T332S/ V37A/ V362I/ WT ΔT371 A364V pr41G EC₅₀ EC₅₀ EC₅₀ EC₅₀ Ex# Structure (μM) (μM) (μM) (μM)   1

0.003 0.017 0.011 0.014   2

3.000 — 1.941 3.000   3

0.009 — 2.218 2.167   4

B B B B   5

0.003 0.015 0.015 0.015   6

A A B A   7

0.002 0.192 0.095 0.192   8

A A A A   9

0.002 — 0.035 0.014  10

0.002 — 0.027 0.006  11

0.002 — 0.018 0.008  12

A A A A  13

0.002 0.007 0.024 0.007  14

0.027 B B B  15

0.004 0.390 2.376 0.390  16

0.005 0.030 0.032 0.030  17

0.003 0.015 B 0.015  18

0.002 A 0.011 0.014  19

0.003 0.047 0.036 0.047  20

A A A A  21

A B A B  22

0.005 0.015 0.008 0.015  23

0.002 A B 0.006  24

0.002 0.012 0.008 0.012  25

0.002 0.010 0.018 0.010  26

0.005 0.041 0.028 0.041  27

A A A A  28

0.003 0.021 0.021 0.021  29

0.005 0.021 0.005 0.021  30

A A A A  31

0.002 0.009 0.006 0.009  32

A A A A  33

0.005 A 0.016 A  34

0.002 0.013 A 0.013  35

0.007 B 0.024 B  36

0.003 0.011 0.005 0.011  37

A B B B  38

0.002 0.232 0.029 0.232  39

0.014 B A B  40

0.004 0.233 0.271 0.233  A1

0.005 0.026 0.009 0.026  A2

0.001 0.008 0.014 0.008  A3

0.004 0.005 0.011 0.005  A4

0.004 0.006 0.026 0.006  A5

0.002 0.003 0.006 0.003  A6

0.002 0.008 0.005 0.008  A7

0.003 0.005 0.477 0.005  A8

0.004 0.023 0.176 0.023  A9

0.003 0.012 0.300 0.012 A11

0.011 0.017 0.281 0.017 A12

0.002 0.059 0.069 0.059 A13

0.002 0.027 0.831 0.027 A14

0.005 0.110 0.114 0.110 A15

0.003 0.010 0.794 0.010 A16

0.010 0.068 0.139 0.068 A17

0.003 0.015 3.000 0.015 A18

0.008 0.027 0.192 0.027 A19

0.003 0.020 B 0.020 A20

A A A A A21

0.003 0.018 0.017 0.018 A22

0.004 0.013 0.027 0.013 A23

0.007 B 0.333 0.193 A24

0.001 3.000 0.007 3.000 A25

A A A A A26

0.005 B B B A27

0.022 B B 3.000 A28

A 0.068 B 0.068 A29

0.005 0.003 0.004 0.003 A30

0.013 0.223 3.000 0.223 A31

0.003 B 3.000 B A32

0.006 B B 0.419 A33

0.006 B 1.787 0.419 A34

0.002 B 0.064 0.096 A35

A 3.000 0.233 B

In Table 3 below, two compounds corresponding to two embodiments of theinvention (Examples 25 and A3) were tested and compared with two other(comparative) compounds outside the scope thereof. Each compound wasassessed for EC₅₀(WT) or EC90 values (see identified strains below,including the T332S/V362I/pr R41G triple mutant):

TABLE 3 T332S/ delV370/ V362I/ WT T371A A364V prR41G EC₅₀ EC₉₀ EC₉₀ EC₉₀Ex # (uM) (uM) (uM) (uM) 25

0.002 0.002 0.041 0.021 A3

0.004 0.015 0.166 0.017 Com- parative

0.003 2.418 0.228 2.418 Com- parative

0.002 1.464 0.340 1.464

As can be deduced from Table 3, the two identified compounds accordingto the invention had better EC90 values versus the comparativecompounds, when tested against the specified mutant strains identifiedabove.

The foregoing description is merely illustrative and should not beunderstood to limit the scope or underlying principles of the inventionin any way. Indeed, various modifications of the invention, in additionto those shown and described herein, will become apparent to thoseskilled in the art from the following examples and the foregoingdescription. Such modifications are also intended to fall within thescope of the appended claims.

What is claimed is:
 1. A pharmaceutical composition comprising thecompound

or salt thereof and further comprising one or more pharmaceuticallyacceptable carriers, excipients, and/or diluents.
 2. A compositionaccording to claim 1 suitable for administration orally, or foradministration parenterally.
 3. A method of treating HIV infection in amammal comprising administering to said mammal a compound or salt ofclaim 1 wherein said administration is oral.
 4. A method of treating HIVinfection in a mammal comprising administering to said mammal a compoundor salt of claim 1 wherein said administration is parenterally.
 5. Themethod of claim 3 wherein said method further comprises administrationof one or more other agents useful for treatment of HIV infection in amammal.
 6. The method of claim 5 wherein said other agent isS/GSK1265744.
 7. The method of claim 4 wherein said method furthercomprises administration of at least one other agent useful fortreatment of HIV infection in a mammal.
 8. The method of claim 7 whereinsaid other agent is S/GSK1265744.